The Experts below are selected from a list of 96 Experts worldwide ranked by ideXlab platform
John J. O'shea - One of the best experts on this subject based on the ideXlab platform.
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analyses of 10 patients and outcomes of stem cell transplantation Janus kinase 3 (JAK3) Deficiency: clinical, immunologic, and molecular
2013Co-Authors: Rebecca H. Buckley, Joseph L. Roberts, Andrea Lengi, Stephanie M. Brown, Min Chen, Yong-jie Zhou, John J. O'sheaAbstract:AbstractWe found 10 individuals from 7 unrelated families among 170 severe combined immunoDeficiency (SCID) patients who exhibited 9 different Janus kinase 3 (JAK3) mutations. These included 3 missense and 2 nonsense mutations, 1 insertion, and 3 deletions. With the exception of one individual with persistence of transplacentally-transferred maternal lymphocytes, all infants presented with a T - B + NK - phenotype. The patient mutations all resulted in abnormal B cell JAK3-dependent IL-2-induced STAT5 phosphorylation. Additional analyses of mutations permitting protein expression revealed the N-terminal JH7 (Del58A) and JH6 (D169E) domain mutations each inhibited receptor binding and catalytic activity, while the G589S JH2 mutation abrogated kinase activity but did not affect c association. Nine of the 10 patients are currently alive from between 4 years and 18 years following stem cell transplantation, with all exhibiting normal T cell function. Reconstitution of antibody function was noted in only 3 patients. NK function was severely depressed at presentation in the 4 patients studied while post-transplant the only individuals with normal NK lytic activity were patients 1 and 5. Hence, bone marrow transplantation is an effective means for reconstitution of T cell immunity in this defect but is less successful for restoration of B cell and NK cell functions.From bloodjournal.hematologylibrary.org by guest on June 3, 2013. For personal use only.
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JAK3-Independent Trafficking of the Common γ Chain Receptor Subunit: Chaperone Function of Jaks Revisited
Molecular and cellular biology, 2004Co-Authors: Sigrun R. Hofmann, Yong-jie Zhou, Albert Q. Lam, Stephan Frank, Haydeé L. Ramos, Yuka Kanno, Davide Agnello, Richard J. Youle, John J. O'sheaAbstract:The Janus family of protein tyrosine kinases (Jaks) is a small family consisting of Jak1, Jak2, JAK3, and Tyk2 (11, 15, 19, 23). These kinases are structurally unique in possessing a carboxy-terminal kinase domain, along with a pseudokinase domain, which gave the Jaks their name. The pseudokinase domain, as its name implies, lacks catalytic activity but has essential regulatory functions (4, 29). Jaks also have an SH2-like domain, but the ability of this region to bind phosphotyrosine has not been established. The amino terminus of Jaks comprises a band-four-point-one, ezrin, radixin, moesin (FERM) domain, which is critical for binding cytokine receptors (7, 8, 23). Many lines of evidence ranging from mutant cell lines to knockout mice and patients with immunoDeficiency indicate that Jaks are essential for signaling via type I and type II cytokine receptors (6, 14, 15, 23). In addition, it has long been appreciated that for some receptors, Jaks also appear to be required for membrane localization of the cognate receptor. That is, the earliest study showing that a Jak, in this case Tyk2, is essential for signaling via alpha/beta interferon (IFN-α/β) also demonstrated that IFN-α receptor subunit 1 (IFNAR1) was poorly expressed on the surface of cells lacking this kinase (35). More recently, it has been reported that Tyk2 impedes constitutive endocytosis and degradation of IFNAR1 (25). This internalization of IFNAR1 is dependent upon the intracellular membrane proximal region (amino acids 480 to 520), since deletion of this region allowed stable surface expression (25). The requirement for Tyk2 for membrane localization of this receptor is most evident in human fibrosarcoma cell lines lacking Tyk2 (7, 26). However, in mice made deficient in Tyk2 by gene targeting, it has been reported that receptor expression is normal (12), although the apparent differences between humans and mice may reflect technical problems related to anti-receptor antibodies. Tyk2 has also been reported to enhance the surface expression of interleukin-10R2 (IL-10R2) (25). Plasma membrane localization of the erythropoietin receptor (EpoR) also requires expression of its cognate Jak, Jak2, although the mechanism appears to be different. EpoR/Jak2 association occurs in the endoplasmic reticulum (ER) and the Jak has chaperone function; in the absence of Jak2, the EpoR fails to traffic to the plasma membrane efficiently. A requirement for Jak has also been reported for the localization of the Oncostatin M receptor to the plasma membrane, although the underlying mechanism has not been elucidated (24). In summary, data from several systems strongly argue for the importance of Jaks in regulating cytokine receptors. However, there are cell- and species-specific differences in this requirement, and a common mechanism for controlling membrane expression has not yet been identified. Unlike other cytokine receptors and Jaks, the common γ chain (γc) and JAK3 have limited distribution, both being predominantly expressed in cells of the hematopoietic lineage (3, 13). Also in contrast to other cytokine receptor-Jak pairs is the selective association of γc and JAK3. To the best of our knowledge, γc interacts exclusively with JAK3 and the converse is also the case. This contention is also supported by genetic data; the phenotype of severe combined immunoDeficiency (SCID) associated with γc Deficiency is identical to that of JAK3 Deficiency (18, 28). These properties are very useful if one wishes to examine whether JAK3 regulates the membrane expression of γc or vice versa. Moreover, mouse and human cells lacking γc, JAK3, or both are available. We therefore set out to determine whether JAK3 and γc regulate each other's subcellular localization. In the present study, by using fluorescent fusion proteins and live cell imaging, we show that, in contrast to the IL-2Rα subunit, γc localized inefficiently to the plasma membrane and accumulated predominantly in endosomal and lysosomal compartments. Measuring receptor surface expression with flow cytometry shows that JAK3 is not required for the expression of γc at the plasma membrane, and its presence or absence does not influence receptor internalization. Nonetheless, overexpression of JAK3 promotes accumulation of γc at the plasma membrane. Conversely, in the absence of γc, JAK3 localized to the cytosol, and importantly, the entire JAK3 molecule is required for the proper localization of this cytokine receptor-Jak pair. These findings demonstrate that the plasma membrane expression of cytokine receptors is not universally dependent upon the cognate Jak. These findings also have important implications for our understanding of Jak structure and indicate that the requirements for proper subcellular localization of Jaks are surprisingly stringent.
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Janus kinase 3 (JAK3) Deficiency: clinical, immunologic, and molecular analyses of 10 patients and outcomes of stem cell transplantation.
Blood, 2003Co-Authors: Joseph L. Roberts, Andrea Lengi, Stephanie M. Brown, Min Chen, Yong-jie Zhou, John J. O'shea, Rebecca H. BuckleyAbstract:We found 10 individuals from 7 unrelated families among 170 severe combined immunoDeficiency (SCID) patients who exhibited 9 different Janus kinase 3 (JAK3) mutations. These included 3 missense and 2 nonsense mutations, 1 insertion, and 3 deletions. With the exception of 1 individual with persistence of transplacentally transferred maternal lymphocytes, all infants presented with a T–B+NK– phenotype. The patient mutations all resulted in abnormal B-cell Janus kinase 3 (JAK3)–dependent interleukin-2 (IL-2)–induced signal transducer and activator of transcription-5 (STAT5) phosphorylation. Additional analyses of mutations permitting protein expression revealed the N-terminal JH7 (del58A) and JH6 (D169E) domain mutations each inhibited receptor binding and catalytic activity, whereas the G589S JH2 mutation abrogated kinase activity but did not affect γc association. Nine of the 10 patients are currently alive from between 4 years and 18 years following stem cell transplantation, with all exhibiting normal T-cell function. Reconstitution of antibody function was noted in only 3 patients. Natural killer (NK) function was severely depressed at presentation in the 4 patients studied, whereas after transplantation the only individuals with normal NK lytic activity were patients 1 and 5. Hence, bone marrow transplantation is an effective means for reconstitution of T-cell immunity in this defect but is less successful for restoration of B-cell and NK cell functions.
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Molecular and biochemical characterization of JAK3 Deficiency in a patient with severe combined immunoDeficiency over 20 years after bone marrow transplantation: Implications for treatment
British journal of haematology, 1998Co-Authors: Fabio Bozzi, Richard Fabian Schumacher, John J. O'shea, Gérard Lefranc, Anna Villa, Raffaele Badolato, Georges Khalil, Jacques Loiselet, Susanna Bresciani, Paolo VezzoniAbstract:Summary. Severe combined immunoDeficiency (SCID) comprises a heterogenous group of disorders that are fatal unless treated by bone marrow transplantation (BMT). The most common form of SCID (T−B+ SCID) is due to mutations of either the common gamma chain (γc) or of γc-coupled JAK3 kinase. We report an unusual JAK3 defect in a female who was successfully treated >20 years ago with a BMT using her HLA-identical father as the donor. Persistence of genetically and biochemically defective autologous B cells, associated with reconstitution of cellular and humoral immunity, suggests that integrity of the γc-JAK3 signalling pathway is not strictly required for immunoglobulin production.
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In vitro correction of JAK3-deficient severe combined immunoDeficiency by retroviral-mediated gene transduction.
The Journal of experimental medicine, 1996Co-Authors: Fabio Candotti, Luigi D. Notarangelo, John J. O'shea, Scott A. Oakes, J A Johnston, R M BlaeseAbstract:Mutations affecting the expression of the Janus family kinase JAK3 were recently shown to be responsible for autosomal recessive severe combined immunoDeficiency (SCID). JAK3-deficient patients present with a clinical phenotype virtually indistinguishable from boys affected by X-linked SCID, a disease caused by genetic defects of the common gamma chain (gamma c) that is a shared component of the receptors for IL-2, IL-4, IL-7, IL-9, and IL-15. The specific interaction of JAK3 and gamma c represents the biochemical basis for the similarities between these two immunodeficiencies. Both forms of SCID are characterized by recurrent, severe infections leading to death in infancy unless successfully treated by allogeneic bone marrow transplantation. Because of the potentially lethal complications associated with allogeneic bone marrow transplantation and the frequent lack of suitable marrow donors, the development of alternative forms of therapy is highly desirable. To this end, we investigated a retroviral-mediated gene correction approach for JAK3-Deficiency. A vector carrying a copy of JAK3 cDNA was constructed and used to transduce B cell lines derived from patients with JAK3-deficient SCID. We demonstrate restoration of JAK3 expression and phosphorylation upon IL-2 and IL-4 stimulation. Furthermore, patients' cells transduced with JAK3 acquired the ability to proliferate normally in response to IL-2. These data indicate that the biological defects of JAK3-deficient cells can be efficiently corrected in vitro by retroviral-mediated gene transfer, thus providing the basis for future investigation of gene therapy as treatment for JAK3-deficient SCID.
Rebecca H. Buckley - One of the best experts on this subject based on the ideXlab platform.
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analyses of 10 patients and outcomes of stem cell transplantation Janus kinase 3 (JAK3) Deficiency: clinical, immunologic, and molecular
2013Co-Authors: Rebecca H. Buckley, Joseph L. Roberts, Andrea Lengi, Stephanie M. Brown, Min Chen, Yong-jie Zhou, John J. O'sheaAbstract:AbstractWe found 10 individuals from 7 unrelated families among 170 severe combined immunoDeficiency (SCID) patients who exhibited 9 different Janus kinase 3 (JAK3) mutations. These included 3 missense and 2 nonsense mutations, 1 insertion, and 3 deletions. With the exception of one individual with persistence of transplacentally-transferred maternal lymphocytes, all infants presented with a T - B + NK - phenotype. The patient mutations all resulted in abnormal B cell JAK3-dependent IL-2-induced STAT5 phosphorylation. Additional analyses of mutations permitting protein expression revealed the N-terminal JH7 (Del58A) and JH6 (D169E) domain mutations each inhibited receptor binding and catalytic activity, while the G589S JH2 mutation abrogated kinase activity but did not affect c association. Nine of the 10 patients are currently alive from between 4 years and 18 years following stem cell transplantation, with all exhibiting normal T cell function. Reconstitution of antibody function was noted in only 3 patients. NK function was severely depressed at presentation in the 4 patients studied while post-transplant the only individuals with normal NK lytic activity were patients 1 and 5. Hence, bone marrow transplantation is an effective means for reconstitution of T cell immunity in this defect but is less successful for restoration of B cell and NK cell functions.From bloodjournal.hematologylibrary.org by guest on June 3, 2013. For personal use only.
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Janus kinase 3 (JAK3) Deficiency: clinical, immunologic, and molecular analyses of 10 patients and outcomes of stem cell transplantation.
Blood, 2003Co-Authors: Joseph L. Roberts, Andrea Lengi, Stephanie M. Brown, Min Chen, Yong-jie Zhou, John J. O'shea, Rebecca H. BuckleyAbstract:We found 10 individuals from 7 unrelated families among 170 severe combined immunoDeficiency (SCID) patients who exhibited 9 different Janus kinase 3 (JAK3) mutations. These included 3 missense and 2 nonsense mutations, 1 insertion, and 3 deletions. With the exception of 1 individual with persistence of transplacentally transferred maternal lymphocytes, all infants presented with a T–B+NK– phenotype. The patient mutations all resulted in abnormal B-cell Janus kinase 3 (JAK3)–dependent interleukin-2 (IL-2)–induced signal transducer and activator of transcription-5 (STAT5) phosphorylation. Additional analyses of mutations permitting protein expression revealed the N-terminal JH7 (del58A) and JH6 (D169E) domain mutations each inhibited receptor binding and catalytic activity, whereas the G589S JH2 mutation abrogated kinase activity but did not affect γc association. Nine of the 10 patients are currently alive from between 4 years and 18 years following stem cell transplantation, with all exhibiting normal T-cell function. Reconstitution of antibody function was noted in only 3 patients. Natural killer (NK) function was severely depressed at presentation in the 4 patients studied, whereas after transplantation the only individuals with normal NK lytic activity were patients 1 and 5. Hence, bone marrow transplantation is an effective means for reconstitution of T-cell immunity in this defect but is less successful for restoration of B-cell and NK cell functions.
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Human severe combined immunoDeficiency: genetic, phenotypic, and functional diversity in one hundred eight infants.
The Journal of pediatrics, 1997Co-Authors: Rebecca H. Buckley, Richard I. Schiff, Sherrie E. Schiff, M. Louise Markert, Larry W. Williams, Terry O. Harville, Joseph L. Roberts, Jennifer M. PuckAbstract:Abstract Objective: To determine the relative frequencies of the different genetic forms of severe combined immunoDeficiency (SCID) and whether there are distinctive characteristics of the particular genotypes. Study design: The demographic, genetic, and immunologic features of 108 infants with SCID who were treated consecutively at Duke University Medical Center were analyzed. Results: Eighty-nine subjects were boys and 19 were girls; there were 84 white infants, 16 black infants, and 8 Hispanic infants. Forty-nine had X-linked SCID with mutations of common cytokine receptor gamma chain (γ c ) , 16 had adenosine deaminase (ADA) Deficiency, 8 had Janus kinase 3 (JAK3) Deficiency, 21 had unknown autosomal recessive mutations, 1 had reticular dysgenesis, 1 had cartilage hair hypoplasia, and 12 (all boys) had SCID of undetermined type. Deficiency of ADA caused the most profound lymphopenia; γ c or JAK3 Deficiency resulted in the most B cells and fewest natural killer (NK) cells; NK cells and function were highest in autosomal recessive and unknown types of SCID. Conclusions: Different SCID genotypes are associated with distinctive lymphocyte characteristics. The presence of NK function in ADA-deficient, autosomal recessive, and unknown type SCIDs, and low NK function in a majority of γ c and JAK3 SCIDs indicates that some molecular lesions affect T, B, and NK cells (γ c and JAK3), others primarily T cells (ADA Deficiency), and others just T and B cells. (J Pediatr 1997;130:378-87)
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HUMAN SEVERE COMBINED IMMUNODeficiency (SCID): GENETIC, PHENOTYPIC AND FUNCTIONAL DIVERSITY IN 95 INFANTS. • 41
Pediatric Research, 1996Co-Authors: Rebecca H. Buckley, Richard I. Schiff, Sherrie E. Schiff, M. Louise Markert, Larry W. Williams, Terry O. Harville, Joseph L. Roberts, Jennifer M. PuckAbstract:Severe combined immunoDeficiency (SCID) is a rare, fatal syndrome of diverse genetic etiology characterized by profound deficiencies of T and B cell function. In the U.S., it affects primarily male infants. Some are due to mutations in the genes encoding adenosine deaminase (ADA), the common gamma chain (Yc) of the IL-2, IL-4, IL-7, IL-9, and IL-15 receptors, or Janus kinase 3 (JAK3). However, for a significant proportion of SCID's, the molecular basis remains unknown. In an effort to identify unique phenotypic and/or functional features that could help identify fundamental causes, we analyzed the clinical, genetic and immunologic features of 95 SCID infants who presented consecutively to this institution between August of 1965 and December of 1995. Eighty (84%) were male and 15 (16%) female; 74 (78%) were white, 13 (14%) black, and 8 (8%) Hispanic. The 95 infants fit 7 categories: 16 (17%) were ADA deficient; 6 (6.3%) had JAK3 Deficiency; 16 (17%) had autosomal recessive inheritance of unknown molecular cause; 1 (1.2%) had reticular dysgenesis; 1 (1.2%) had cartilage hair hypoplasia; 38 (40%) had X-linked SCID; and 17 (18%) (all males) had SCID of unknown type. Numbers of total lymphocytes and the various lymphocyte phenotypes distinguished the groups: ADA deficient SCID's had the lowest numbers of total lymphocytes; B cells were present in highest number and NK cells in lowest number in X-linked and JAK3-deficient SCIDs. Mean numbers of NK cells were highest in autosomal recessive and unknown types of SCID. All types were profoundly deficient in T cells. NK function was tested in 60 and found to be normal in 25. Although the family history was positive in only 34% of X-linked cases, it is likely that as many as two-thirds of the boys with SCID have mutations in Yc. The presence of NK function in ADA deficient, autosomal recessive, and some SCID's of unknown type, and abnormally low NK numbers and function in a majority of X-linked and JAK3-deficient SCID's implies that some molecular lesions affect T, B and NK cells (Yc and JAK3 mutations), but that others affect only T cells (ADA Deficiency), and others T and B cells (possible recombinase abnormalities). The findings in this group of SCID's, the largest reported in the U.S., argue against a common lineage for T and NK cells and may provide clues as to the molecular bases of SCID in the 35 (37%) infants who did not have X-linked SCID, JAK3 Deficiency or ADA Deficiency.
Jennifer M. Puck - One of the best experts on this subject based on the ideXlab platform.
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Perspectives of gene therapy for primary immunodeficiencies.
Current opinion in allergy and clinical immunology, 2004Co-Authors: Javier Chinen, Jennifer M. PuckAbstract:PURPOSE OF REVIEW Standard therapies for patients with severe primary immunodeficiencies include bone marrow transplantation and, for adenosine deaminase Deficiency, enzyme replacement. In the last decade, gene therapy has been developed as an alternative for these conditions. We summarize the recent advances in gene therapy for primary immunodeficiencies and discuss the unexpected occurrence of leukemia in a gene therapy trial for X-linked severe combined immunoDeficiency. RECENT FINDINGS Eight of 10 infants with X-linked severe combined immunoDeficiency who received autologous hematopoietic stem cells transduced with a retroviral vector carrying the IL2RG complementary DNA achieved immune reconstitution. However, the two youngest patients developed leukemic expansions of gene-corrected cells. The first case had proliferation of a gamma delta T cell clone, and the second case had three alpha beta T cell clones derived from a single transduced progenitor. Leukemic cells in both patients aberrantly expressed the LIM domain only-2 transcription factor due to retroviral vector insertions in this locus. After receiving anti-leukemic treatment one patient achieved a lasting remission, but the other relapsed. Four adenosine deaminase deficient severe combined immunoDeficiency patients also developed functional immunity after receiving autologous hematopoietic stem cells transduced with the adenosine deaminase gene complementary DNA following submyeloablative chemotherapy. Chronic granulomatous disease, Wiskott-Aldrich syndrome, JAK3 Deficiency and RAG2 Deficiency are other immunodeficiencies being studied as candidates for gene therapy. SUMMARY Gene therapy is a promising therapeutic option for some primary immunodeficiencies, especially when cells expressing the correct gene have a selective advantage. More clinical trials with closer patient monitoring are under way to define which patients may benefit from this approach, and strategies are being developed to understand and ultimately reduce the risk of leukemia secondary to retroviral vector insertion.
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Human severe combined immunoDeficiency: genetic, phenotypic, and functional diversity in one hundred eight infants.
The Journal of pediatrics, 1997Co-Authors: Rebecca H. Buckley, Richard I. Schiff, Sherrie E. Schiff, M. Louise Markert, Larry W. Williams, Terry O. Harville, Joseph L. Roberts, Jennifer M. PuckAbstract:Abstract Objective: To determine the relative frequencies of the different genetic forms of severe combined immunoDeficiency (SCID) and whether there are distinctive characteristics of the particular genotypes. Study design: The demographic, genetic, and immunologic features of 108 infants with SCID who were treated consecutively at Duke University Medical Center were analyzed. Results: Eighty-nine subjects were boys and 19 were girls; there were 84 white infants, 16 black infants, and 8 Hispanic infants. Forty-nine had X-linked SCID with mutations of common cytokine receptor gamma chain (γ c ) , 16 had adenosine deaminase (ADA) Deficiency, 8 had Janus kinase 3 (JAK3) Deficiency, 21 had unknown autosomal recessive mutations, 1 had reticular dysgenesis, 1 had cartilage hair hypoplasia, and 12 (all boys) had SCID of undetermined type. Deficiency of ADA caused the most profound lymphopenia; γ c or JAK3 Deficiency resulted in the most B cells and fewest natural killer (NK) cells; NK cells and function were highest in autosomal recessive and unknown types of SCID. Conclusions: Different SCID genotypes are associated with distinctive lymphocyte characteristics. The presence of NK function in ADA-deficient, autosomal recessive, and unknown type SCIDs, and low NK function in a majority of γ c and JAK3 SCIDs indicates that some molecular lesions affect T, B, and NK cells (γ c and JAK3), others primarily T cells (ADA Deficiency), and others just T and B cells. (J Pediatr 1997;130:378-87)
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HUMAN SEVERE COMBINED IMMUNODeficiency (SCID): GENETIC, PHENOTYPIC AND FUNCTIONAL DIVERSITY IN 95 INFANTS. • 41
Pediatric Research, 1996Co-Authors: Rebecca H. Buckley, Richard I. Schiff, Sherrie E. Schiff, M. Louise Markert, Larry W. Williams, Terry O. Harville, Joseph L. Roberts, Jennifer M. PuckAbstract:Severe combined immunoDeficiency (SCID) is a rare, fatal syndrome of diverse genetic etiology characterized by profound deficiencies of T and B cell function. In the U.S., it affects primarily male infants. Some are due to mutations in the genes encoding adenosine deaminase (ADA), the common gamma chain (Yc) of the IL-2, IL-4, IL-7, IL-9, and IL-15 receptors, or Janus kinase 3 (JAK3). However, for a significant proportion of SCID's, the molecular basis remains unknown. In an effort to identify unique phenotypic and/or functional features that could help identify fundamental causes, we analyzed the clinical, genetic and immunologic features of 95 SCID infants who presented consecutively to this institution between August of 1965 and December of 1995. Eighty (84%) were male and 15 (16%) female; 74 (78%) were white, 13 (14%) black, and 8 (8%) Hispanic. The 95 infants fit 7 categories: 16 (17%) were ADA deficient; 6 (6.3%) had JAK3 Deficiency; 16 (17%) had autosomal recessive inheritance of unknown molecular cause; 1 (1.2%) had reticular dysgenesis; 1 (1.2%) had cartilage hair hypoplasia; 38 (40%) had X-linked SCID; and 17 (18%) (all males) had SCID of unknown type. Numbers of total lymphocytes and the various lymphocyte phenotypes distinguished the groups: ADA deficient SCID's had the lowest numbers of total lymphocytes; B cells were present in highest number and NK cells in lowest number in X-linked and JAK3-deficient SCIDs. Mean numbers of NK cells were highest in autosomal recessive and unknown types of SCID. All types were profoundly deficient in T cells. NK function was tested in 60 and found to be normal in 25. Although the family history was positive in only 34% of X-linked cases, it is likely that as many as two-thirds of the boys with SCID have mutations in Yc. The presence of NK function in ADA deficient, autosomal recessive, and some SCID's of unknown type, and abnormally low NK numbers and function in a majority of X-linked and JAK3-deficient SCID's implies that some molecular lesions affect T, B and NK cells (Yc and JAK3 mutations), but that others affect only T cells (ADA Deficiency), and others T and B cells (possible recombinase abnormalities). The findings in this group of SCID's, the largest reported in the U.S., argue against a common lineage for T and NK cells and may provide clues as to the molecular bases of SCID in the 35 (37%) infants who did not have X-linked SCID, JAK3 Deficiency or ADA Deficiency.
Luigi D. Notarangelo - One of the best experts on this subject based on the ideXlab platform.
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Mutations in severe combined immune Deficiency (SCID) due to JAK3 Deficiency
Human mutation, 2001Co-Authors: Luigi D. Notarangelo, Patrizia Mella, Alison L Jones, Geneviève De Saint Basile, Gianfranco Savoldi, Treena Cranston, Mauno Vihinen, Richard Fabian SchumacherAbstract:During the last 10 years, an increasing number of genes have been identified whose abnormalities account for primary immunodeficiencies, with defects in development and/or function of the immune system. Among them is the JAK3,gene, encoding for a tyrosine kinase that is functionally coupled to cytokine receptors which share the common gamma chain. Defects of this gene cause an autosomal recessive form of severe combined immunoDeficiency with almost absent T cells and functionally defective B cells (T-B+ SCID). Herewith, we present molecular information on the first 27 unique mutations identified in the JAK3 gene, including clinical data on all of the 23 affected patients reported so far. A variety of mutations scattered throughout all seven functional domains of the protein, and with different functional effects, have been identified. Availability of a molecular screening test, based on amplification of genomic DNA, facilitates the diagnostic approach, and has permitted recognition that JAK3 Deficiency may also be associated with atypical clinical and immunological features. Development of a structural model of the JAK3 kinase domain has allowed characterization of the functional effects of the various mutations. Most importantly, molecular analysis at the JAK3 locus results in improved genetic counseling, allows early prenatal diagnosis, and prompts appropriate treatment (currently based on hematopoietic stem cell transplantation) in affected families. Hum Mutat 18:255-263, 2001. (C) 2001 Wiley-Liss,Inc. (Less)
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Of genes and phenotypes: the immunological and molecular spectrum of combined immune Deficiency. Defects of the gamma(c)-JAK3 signaling pathway as a model
Immunological reviews, 2000Co-Authors: Luigi D. Notarangelo, Patrizia Mella, Gianfranco Savoldi, Silvia Giliani, Cinzia Mazza, Carmen Rodriguez-perez, Evelina Mazzolari, Maurilia Fiorini, Marzia Duse, Alessandro PlebaniAbstract:Cytokines play a major role in lymphoid development. Defects of the common gamma chain (gamma(c)) or of the JAK3 protein in humans have been shown to result in a severe combined immune Deficiency (SCID), with a profound defect in T and natural killer (NK)-cell development, whereas B-cell generation is apparently unaffected (T-B+NK-SCID). While extensive molecular and biochemical analysis of these patients has been instrumental in understanding better the biological properties of the gamma(c) and JAK3 protein, an unexpected phenotypic heterogeneity of gamma(c) and JAK3 Deficiency has emerged, indicating the need for appropriate and extensive investigations even in patients with atypical presentations. At the same time, characterization of the defects has been instrumental in the development of novel therapeutic approaches, from in utero hematopoietic stem cell transplantation to gene therapy.
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JAK3-DEFICIENT SEVERE COMBINED IMMUNODeficiency
Immunology and Allergy Clinics of North America, 2000Co-Authors: Luigi D. Notarangelo, Fabio CandottiAbstract:Severe combined immunoDeficiency (SCID) comprises a heterogeneous group of genetic disorders, with numerical and functional defects of both T and B lymphocytes. The most common form of SCID in humans, accounting for about 40% to 50% of all cases, is characterized by lack of circulating T (and usually also NK) cells, with a normal or increased number of B lymphocytes (T − B + SCID). 5,47 Most patients with T − B + SCID are men, reflecting the occurrence of X-linked SCID (SCIDX1), a disorder caused by mutations of the IL2RG gene, 31,38,39 encoding for the common gamma chain (γ c ), shared by receptors for interleukin (IL)-2, IL-4, IL-7, IL-9, and IL-15. 25,48 In all these receptors, γ c is associated with the intracellular tyrosine kinase JAK3, whereas the unique receptor transducing subunit (as the β chain in the case of the IL-2 receptor, IL-2Rβ) binds to another JAK kinase, JAK1. 29 JAK1 and JAK3 are strictly required for cytokine-mediated signaling. Mutations of the IL2RG gene that affect interaction between the γ c and JAK3 also cause SCID. 42 Based on these data, it was hypothesized and then demonstrated in three unrelated patients, that mutations of the JAK3 gene are responsible for autosomal recessive T − B + SCID in humans. 27,43 Disruption of the JAK3 gene in mice also results in a form of murine SCID , that is indistinguishable by that of γ c −/y mice. 32,36,49 During recent years, additional patients with SCID caused by JAK3 Deficiency have been identified 3,11 ; this has allowed better definition of the clinical and immunologic spectrum of the disease, its biochemical basis, and evaluation of prognostic and therapeutic implications of this disorder. Finally, the potential for gene therapy to correct this disorder has been successfully investigated in the murine knock-out model 7,8 and through in vitro biochemical correction of JAK3-deficient human cell lines. 10
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In vitro correction of JAK3-deficient severe combined immunoDeficiency by retroviral-mediated gene transduction.
The Journal of experimental medicine, 1996Co-Authors: Fabio Candotti, Luigi D. Notarangelo, John J. O'shea, Scott A. Oakes, J A Johnston, R M BlaeseAbstract:Mutations affecting the expression of the Janus family kinase JAK3 were recently shown to be responsible for autosomal recessive severe combined immunoDeficiency (SCID). JAK3-deficient patients present with a clinical phenotype virtually indistinguishable from boys affected by X-linked SCID, a disease caused by genetic defects of the common gamma chain (gamma c) that is a shared component of the receptors for IL-2, IL-4, IL-7, IL-9, and IL-15. The specific interaction of JAK3 and gamma c represents the biochemical basis for the similarities between these two immunodeficiencies. Both forms of SCID are characterized by recurrent, severe infections leading to death in infancy unless successfully treated by allogeneic bone marrow transplantation. Because of the potentially lethal complications associated with allogeneic bone marrow transplantation and the frequent lack of suitable marrow donors, the development of alternative forms of therapy is highly desirable. To this end, we investigated a retroviral-mediated gene correction approach for JAK3-Deficiency. A vector carrying a copy of JAK3 cDNA was constructed and used to transduce B cell lines derived from patients with JAK3-deficient SCID. We demonstrate restoration of JAK3 expression and phosphorylation upon IL-2 and IL-4 stimulation. Furthermore, patients' cells transduced with JAK3 acquired the ability to proliferate normally in response to IL-2. These data indicate that the biological defects of JAK3-deficient cells can be efficiently corrected in vitro by retroviral-mediated gene transfer, thus providing the basis for future investigation of gene therapy as treatment for JAK3-deficient SCID.
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In Vitro Correction ofJAK3-deficient Severe Combined ImmunoDeficiency by Retroviral-mediated
1996Co-Authors: Fabio Candotti, Luigi D. Notarangelo, John J. O'shea, Scott A. Oakes, R M BlaeseAbstract:Summary Mutations affecting the expression of the Janus family kinase JAK3 were recently shown to be responsible for autosomal recessive severe combined immunoDeficiency (SCID). JAK3-deficient patients present with a clinical phenotype virtually indistinguishable from boys affected by X-linked SCID, a disease caused by genetic defects of the common gamma chain (%) that is a shared component of the receptors for IL-2, IL-4, IL-7, IL-9, and IL-15. The specific interaction of JAK3 and ~/c represents the biochemical basis for the similarities between these two immunodeficiencies. forms of SCID are characterized by recurrent, severe infections leading to death in infancy unless successfully treated by allogeneic bone marrow transplantation. Because of the potentially lethal complications associated with allogeneic bone marrow transplantation and the frequent lack of suitable marrow donors, the development of alternative forms of therapy is highly desirable. To this end, we investigated a retroviral-mediated gene correction approach for JAK3-Deficiency. A vector carrying a copy ofJAK3 cDNA was constructed and used to transduce B cell lines derived from patients with JAK3-deficient SCID. We demonstrate restoration of JAK3 expression and phosphorylation upon IL-2 and IL-4 stimulation. Furthermore, patients' cells transduced with JAK3 acquired the ability to proliferate normally in response to IL-2. These data indicate that the biological defects of JAK3-deficient cells can be efficiently corrected in vitro by retroviral-mediated gene transfer, thus providing the basis for future investigation of gene therapy as treatment for JAK3-deficient SCID.
Joseph L. Roberts - One of the best experts on this subject based on the ideXlab platform.
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analyses of 10 patients and outcomes of stem cell transplantation Janus kinase 3 (JAK3) Deficiency: clinical, immunologic, and molecular
2013Co-Authors: Rebecca H. Buckley, Joseph L. Roberts, Andrea Lengi, Stephanie M. Brown, Min Chen, Yong-jie Zhou, John J. O'sheaAbstract:AbstractWe found 10 individuals from 7 unrelated families among 170 severe combined immunoDeficiency (SCID) patients who exhibited 9 different Janus kinase 3 (JAK3) mutations. These included 3 missense and 2 nonsense mutations, 1 insertion, and 3 deletions. With the exception of one individual with persistence of transplacentally-transferred maternal lymphocytes, all infants presented with a T - B + NK - phenotype. The patient mutations all resulted in abnormal B cell JAK3-dependent IL-2-induced STAT5 phosphorylation. Additional analyses of mutations permitting protein expression revealed the N-terminal JH7 (Del58A) and JH6 (D169E) domain mutations each inhibited receptor binding and catalytic activity, while the G589S JH2 mutation abrogated kinase activity but did not affect c association. Nine of the 10 patients are currently alive from between 4 years and 18 years following stem cell transplantation, with all exhibiting normal T cell function. Reconstitution of antibody function was noted in only 3 patients. NK function was severely depressed at presentation in the 4 patients studied while post-transplant the only individuals with normal NK lytic activity were patients 1 and 5. Hence, bone marrow transplantation is an effective means for reconstitution of T cell immunity in this defect but is less successful for restoration of B cell and NK cell functions.From bloodjournal.hematologylibrary.org by guest on June 3, 2013. For personal use only.
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Janus kinase 3 (JAK3) Deficiency: clinical, immunologic, and molecular analyses of 10 patients and outcomes of stem cell transplantation.
Blood, 2003Co-Authors: Joseph L. Roberts, Andrea Lengi, Stephanie M. Brown, Min Chen, Yong-jie Zhou, John J. O'shea, Rebecca H. BuckleyAbstract:We found 10 individuals from 7 unrelated families among 170 severe combined immunoDeficiency (SCID) patients who exhibited 9 different Janus kinase 3 (JAK3) mutations. These included 3 missense and 2 nonsense mutations, 1 insertion, and 3 deletions. With the exception of 1 individual with persistence of transplacentally transferred maternal lymphocytes, all infants presented with a T–B+NK– phenotype. The patient mutations all resulted in abnormal B-cell Janus kinase 3 (JAK3)–dependent interleukin-2 (IL-2)–induced signal transducer and activator of transcription-5 (STAT5) phosphorylation. Additional analyses of mutations permitting protein expression revealed the N-terminal JH7 (del58A) and JH6 (D169E) domain mutations each inhibited receptor binding and catalytic activity, whereas the G589S JH2 mutation abrogated kinase activity but did not affect γc association. Nine of the 10 patients are currently alive from between 4 years and 18 years following stem cell transplantation, with all exhibiting normal T-cell function. Reconstitution of antibody function was noted in only 3 patients. Natural killer (NK) function was severely depressed at presentation in the 4 patients studied, whereas after transplantation the only individuals with normal NK lytic activity were patients 1 and 5. Hence, bone marrow transplantation is an effective means for reconstitution of T-cell immunity in this defect but is less successful for restoration of B-cell and NK cell functions.
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Human severe combined immunoDeficiency: genetic, phenotypic, and functional diversity in one hundred eight infants.
The Journal of pediatrics, 1997Co-Authors: Rebecca H. Buckley, Richard I. Schiff, Sherrie E. Schiff, M. Louise Markert, Larry W. Williams, Terry O. Harville, Joseph L. Roberts, Jennifer M. PuckAbstract:Abstract Objective: To determine the relative frequencies of the different genetic forms of severe combined immunoDeficiency (SCID) and whether there are distinctive characteristics of the particular genotypes. Study design: The demographic, genetic, and immunologic features of 108 infants with SCID who were treated consecutively at Duke University Medical Center were analyzed. Results: Eighty-nine subjects were boys and 19 were girls; there were 84 white infants, 16 black infants, and 8 Hispanic infants. Forty-nine had X-linked SCID with mutations of common cytokine receptor gamma chain (γ c ) , 16 had adenosine deaminase (ADA) Deficiency, 8 had Janus kinase 3 (JAK3) Deficiency, 21 had unknown autosomal recessive mutations, 1 had reticular dysgenesis, 1 had cartilage hair hypoplasia, and 12 (all boys) had SCID of undetermined type. Deficiency of ADA caused the most profound lymphopenia; γ c or JAK3 Deficiency resulted in the most B cells and fewest natural killer (NK) cells; NK cells and function were highest in autosomal recessive and unknown types of SCID. Conclusions: Different SCID genotypes are associated with distinctive lymphocyte characteristics. The presence of NK function in ADA-deficient, autosomal recessive, and unknown type SCIDs, and low NK function in a majority of γ c and JAK3 SCIDs indicates that some molecular lesions affect T, B, and NK cells (γ c and JAK3), others primarily T cells (ADA Deficiency), and others just T and B cells. (J Pediatr 1997;130:378-87)
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HUMAN SEVERE COMBINED IMMUNODeficiency (SCID): GENETIC, PHENOTYPIC AND FUNCTIONAL DIVERSITY IN 95 INFANTS. • 41
Pediatric Research, 1996Co-Authors: Rebecca H. Buckley, Richard I. Schiff, Sherrie E. Schiff, M. Louise Markert, Larry W. Williams, Terry O. Harville, Joseph L. Roberts, Jennifer M. PuckAbstract:Severe combined immunoDeficiency (SCID) is a rare, fatal syndrome of diverse genetic etiology characterized by profound deficiencies of T and B cell function. In the U.S., it affects primarily male infants. Some are due to mutations in the genes encoding adenosine deaminase (ADA), the common gamma chain (Yc) of the IL-2, IL-4, IL-7, IL-9, and IL-15 receptors, or Janus kinase 3 (JAK3). However, for a significant proportion of SCID's, the molecular basis remains unknown. In an effort to identify unique phenotypic and/or functional features that could help identify fundamental causes, we analyzed the clinical, genetic and immunologic features of 95 SCID infants who presented consecutively to this institution between August of 1965 and December of 1995. Eighty (84%) were male and 15 (16%) female; 74 (78%) were white, 13 (14%) black, and 8 (8%) Hispanic. The 95 infants fit 7 categories: 16 (17%) were ADA deficient; 6 (6.3%) had JAK3 Deficiency; 16 (17%) had autosomal recessive inheritance of unknown molecular cause; 1 (1.2%) had reticular dysgenesis; 1 (1.2%) had cartilage hair hypoplasia; 38 (40%) had X-linked SCID; and 17 (18%) (all males) had SCID of unknown type. Numbers of total lymphocytes and the various lymphocyte phenotypes distinguished the groups: ADA deficient SCID's had the lowest numbers of total lymphocytes; B cells were present in highest number and NK cells in lowest number in X-linked and JAK3-deficient SCIDs. Mean numbers of NK cells were highest in autosomal recessive and unknown types of SCID. All types were profoundly deficient in T cells. NK function was tested in 60 and found to be normal in 25. Although the family history was positive in only 34% of X-linked cases, it is likely that as many as two-thirds of the boys with SCID have mutations in Yc. The presence of NK function in ADA deficient, autosomal recessive, and some SCID's of unknown type, and abnormally low NK numbers and function in a majority of X-linked and JAK3-deficient SCID's implies that some molecular lesions affect T, B and NK cells (Yc and JAK3 mutations), but that others affect only T cells (ADA Deficiency), and others T and B cells (possible recombinase abnormalities). The findings in this group of SCID's, the largest reported in the U.S., argue against a common lineage for T and NK cells and may provide clues as to the molecular bases of SCID in the 35 (37%) infants who did not have X-linked SCID, JAK3 Deficiency or ADA Deficiency.