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Landon Trost - One of the best experts on this subject based on the ideXlab platform.
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Treatment Patterns and Healthcare Outcomes with Collagenase Clostridium histolyticum vs Surgery in Peyronie's Disease: A Retrospective Claims Database Analysis
Sexual medicine, 2021Co-Authors: Landon Trost, Huan Huang, Xu Han, Chakkarin BurudpakdeeAbstract:Abstract Introduction Treatments for Peyronie's disease (PD) include surgical management and collagenase Clostridium histolyticum (CCH). Aims To evaluate PD treatment trends after CCH approval and compare clinical outcomes in CCH- and surgery-treated cohorts. Methods Patients newly diagnosed with PD between January 2011 and December 2017 were identified in a U.S. claims database. Cohorts initiating treatment with CCH or surgery between January 2014 and June 2017 were included. Patients were continuously enrolled ≥6 months before and ≥12 months after index date. Post-treatment penile complications and analgesic use were compared 1 year after procedure in propensity score-matched cohorts. Main outcome measures The main outcome measures of this study were treatment patterns, penile complications, and analgesic use. Results In the newly diagnosed PD cohort, 1,609 patients received CCH and 1,555 patients had surgery. Overall CCH or surgery treatment rate/year increased from 9.8% in 2014 to 15.5% in 2017, with Conclusion CCH demonstrated fewer complications and less analgesic use than surgery and was used as the initial therapy for PD twice as often as surgery. L Trost, H Huang, X Han, et al. Treatment Patterns and Healthcare Outcomes with Collagenase Clostridium histolyticum vs Surgery in Peyronie's Disease: A Retrospective Claims Database Analysis. Sex Med 2021;XX:XXX–XXX.
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intralesional collagenase Clostridium histolyticum causes meaningful improvement in men with peyronie s disease results of a multi institutional analysis
The Journal of Urology, 2019Co-Authors: Wayne J.g. Hellstrom, Landon Trost, Hoang Minh Tue Nguyen, Laith Alzweri, Amanda S J Chung, Ramon Virasoro, Ashley Tapscott, Matthew Ziegelmann, Martin K. GelbardAbstract:Purpose:In a multi-institutional setting we studied the efficacy and safety outcomes at multiple high volume centers where collagenase Clostridium histolyticum is used to treat Peyronie's disease.M...
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restoration of penile function and patient satisfaction with intralesional collagenase Clostridium histolyticum injection for peyronie s disease
The Journal of Urology, 2016Co-Authors: Matthew J Ziegelmann, Boyd R Viers, Kelly L Mcalvany, George C Bailey, Joshua Savage, Landon TrostAbstract:Purpose: Collagenase Clostridium histolyticum is approved for the treatment of Peyronie’s disease. To date, no post-release study to our knowledge has evaluated patient perceived outcomes and satisfaction. Therefore, we evaluated patient perceived experience with collagenase Clostridium histolyticum injection for Peyronie’s disease in a clinical practice.Materials and Methods: From March 2014 to July 2015, 69 patients underwent 1 to 4 series of collagenase Clostridium histolyticum injections for Peyronie’s disease at our institution. Objective changes in penile curvature as well as patient reported functional outcomes and patient perceived curvature improvements were evaluated.Results: By the time of analysis 31 patients (45%) had completed 4 trials, 47 (68%) completed 3 trials and 59 (86%) completed 2 trials. Patient reported improvements (percentage) in curvature increased with each series (trial 1—14%, trial 2—28%, trial 3—30% and trial 4—37%, p <0.05). Among those completing therapy 57% reported that ...
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Restoration of Penile Function and Patient Satisfaction with Intralesional Collagenase Clostridium histolyticum Injection for Peyronie’s Disease
The Journal of Urology, 2015Co-Authors: Matthew J Ziegelmann, Boyd R Viers, Kelly L Mcalvany, George C Bailey, Joshua Savage, Landon TrostAbstract:Purpose: Collagenase Clostridium histolyticum is approved for the treatment of Peyronie’s disease. To date, no post-release study to our knowledge has evaluated patient perceived outcomes and satisfaction. Therefore, we evaluated patient perceived experience with collagenase Clostridium histolyticum injection for Peyronie’s disease in a clinical practice.Materials and Methods: From March 2014 to July 2015, 69 patients underwent 1 to 4 series of collagenase Clostridium histolyticum injections for Peyronie’s disease at our institution. Objective changes in penile curvature as well as patient reported functional outcomes and patient perceived curvature improvements were evaluated.Results: By the time of analysis 31 patients (45%) had completed 4 trials, 47 (68%) completed 3 trials and 59 (86%) completed 2 trials. Patient reported improvements (percentage) in curvature increased with each series (trial 1—14%, trial 2—28%, trial 3—30% and trial 4—37%, p
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Collagenase Clostridium histolyticum: a novel nonsurgical option for the treatment of Peyronie’s disease
International Journal of Clinical Rheumatology, 2013Co-Authors: Louis A. Aliperti, Landon Trost, Wayne J.g. HellstromAbstract:Commercially available collagenase Clostridium histolyticum (CCH) comprises a mixture of clostridial type-I/II collagenase enzymes, which enzymatically destroy collagen deposits. CCH is currently approved in the USA, EU and Canada for the treatment of Dupuytren’s contracture with a palpable cord in the hands of adult patients. Intralesional injection of CCH has demonstrated safety and efficacy in the treatment of Peyronie’s disease (PD) in Phase I–III clinical trials. Improvement of penile curvature deformity and psychological symptoms (degree of patient bother associated with PD) has been achieved among patients with PD. CCH is safe and well tolerated, with adverse events localized to the injection site. This article reviews the potential mechanism of action and summarizes the clinical experience with CCH in patients with PD.
Faysal A. Yafi - One of the best experts on this subject based on the ideXlab platform.
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Management of Peyronie’s disease with collagenase Clostridium histolyticum in the acute phase
World Journal of Urology, 2020Co-Authors: Farouk M. El-khatib, Maxwell Towe, Faysal A. YafiAbstract:Introduction Peyronie’s disease (PD) can affect men of all ages and is associated with penile pain as well as curvature and erectile dysfunction. Treatment modalities for PD include conservative, less invasive and surgical treatments. Choice of treatment depends on the disease’s phase. Injection of collagenase Clostridium histolyticum (CCH) is only the Food and Drug Administration approved treatment for the management of the chronic phase of the disease. Only a few limited studies have evaluated its potential benefits during the acute phase. Aim To evaluate the current evidence on the use of injectable CCH during the acute or active phase of PD. Methods We performed a PubMed database search for articles published between 2015 and 2018 that investigated the use of CCH for the management of the acute phase of PD. Search keywords included “Peyronie’s disease”, “collagenase Clostridium histolyticum ”, and “acute phase”. Main outcome measures Changes in penile curvature after treatment with injectable CCH. Results Mean curvature decrease ranged between 15.8° and 22.6° corresponding to 27.4–37.4% decrease from baseline after 2.5–4 cycles, corresponding to 5–8 injections, of CCH treatment in patients during the acute phase of PD. Conclusions Intralesional CCH injection therapy during the acute phase of PD can be both safe and effective and clinically significant improvements in penile curvature may be achieved. Larger multi-institutional studies are, however, still needed to confirm these results and validate this additional indication for CCH.
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management of peyronie s disease with collagenase Clostridium histolyticum in the acute phase
World Journal of Urology, 2020Co-Authors: Farouk M Elkhatib, Maxwell Towe, Faysal A. YafiAbstract:Peyronie’s disease (PD) can affect men of all ages and is associated with penile pain as well as curvature and erectile dysfunction. Treatment modalities for PD include conservative, less invasive and surgical treatments. Choice of treatment depends on the disease’s phase. Injection of collagenase Clostridium histolyticum (CCH) is only the Food and Drug Administration approved treatment for the management of the chronic phase of the disease. Only a few limited studies have evaluated its potential benefits during the acute phase. To evaluate the current evidence on the use of injectable CCH during the acute or active phase of PD. We performed a PubMed database search for articles published between 2015 and 2018 that investigated the use of CCH for the management of the acute phase of PD. Search keywords included “Peyronie’s disease”, “collagenase Clostridium histolyticum”, and “acute phase”. Changes in penile curvature after treatment with injectable CCH. Mean curvature decrease ranged between 15.8° and 22.6° corresponding to 27.4–37.4% decrease from baseline after 2.5–4 cycles, corresponding to 5–8 injections, of CCH treatment in patients during the acute phase of PD. Intralesional CCH injection therapy during the acute phase of PD can be both safe and effective and clinically significant improvements in penile curvature may be achieved. Larger multi-institutional studies are, however, still needed to confirm these results and validate this additional indication for CCH.
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Management of Peyronie’s disease with collagenase Clostridium histolyticum in the acute phase
World journal of urology, 2019Co-Authors: Farouk M. El-khatib, Maxwell Towe, Faysal A. YafiAbstract:Peyronie’s disease (PD) can affect men of all ages and is associated with penile pain as well as curvature and erectile dysfunction. Treatment modalities for PD include conservative, less invasive and surgical treatments. Choice of treatment depends on the disease’s phase. Injection of collagenase Clostridium histolyticum (CCH) is only the Food and Drug Administration approved treatment for the management of the chronic phase of the disease. Only a few limited studies have evaluated its potential benefits during the acute phase. To evaluate the current evidence on the use of injectable CCH during the acute or active phase of PD. We performed a PubMed database search for articles published between 2015 and 2018 that investigated the use of CCH for the management of the acute phase of PD. Search keywords included “Peyronie’s disease”, “collagenase Clostridium histolyticum”, and “acute phase”. Changes in penile curvature after treatment with injectable CCH. Mean curvature decrease ranged between 15.8° and 22.6° corresponding to 27.4–37.4% decrease from baseline after 2.5–4 cycles, corresponding to 5–8 injections, of CCH treatment in patients during the acute phase of PD. Intralesional CCH injection therapy during the acute phase of PD can be both safe and effective and clinically significant improvements in penile curvature may be achieved. Larger multi-institutional studies are, however, still needed to confirm these results and validate this additional indication for CCH.
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MP65-08 INTRALESIONAL INJECTION OF COLLAGENASE Clostridium histolyticum FOR THE MANAGEMENT OF PEYRONIE’S DISEASE WITH INDENTATION OR HOURGLASS DEFORMITY
Journal of Urology, 2019Co-Authors: M.m. Osman, Maxwell Towe, Farouk M. El-khatib, Linda Huynh, Faysal A. YafiAbstract:INTRODUCTION AND OBJECTIVES:Intralesional injection (ILI) of collagenase Clostridium histolyticum (CCH) is the only Food and Drug Administration approved medical treatment for the management of sta...
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Intralesional collagenase Clostridium histolyticum in the management of Peyronie's disease: current best practice.
Therapeutic advances in urology, 2018Co-Authors: Daniel J. Cwikla, Faysal A. YafiAbstract:The use of Clostridium histolyticum collagenase (CCH) has become increasingly widespread for the treatment of Peyronie’s disease (PD) in recent years. Numerous trials have confirmed both its safety...
Claudiu T. Supuran - One of the best experts on this subject based on the ideXlab platform.
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Protease Inhibitors. Part 5. Alkyl/Arylsulfonyl‐ and Arylsulfonylureido‐/Arylureido‐ Glycine Hydroxamate Inhibitors of Clostridium histolyticum Collagenase.
ChemInform, 2010Co-Authors: Andrea Scozzafava, Claudiu T. SupuranAbstract:Abstract Reaction of alkyl/arylsulfonyl halides with glycine afforded a series of derivatives which were first N -benzylated by treatment with benzyl chloride, and then converted to the corresponding hydroxamic acids with hydroxylamine in the presence of carbodiimide derivatives. Other derivatives were obtained by reaction of N -benzyl-glycine with aryl isocyanates, arylsulfonyl isocyanates or benzoyl isothiocyanate, followed by conversion of their COOH group into the CONHOH moiety, as mentioned above. The 90 new compounds reported here were assayed as inhibitors of the Clostridium histolyticum collagenase (EC 3.4.24.3), a zinc enzyme which degrades triple helical regions of native collagen. The prepared hydroxamate derivatives were generally 100–500 times more active than the corresponding carboxylates. In the series of synthesized hydroxamates, substitution patterns leading to the best inhibitors were those involving perfluoroalkylsulfonyl- and substituted-arylsulfonyl moieties, such as pentafluorophenylsulfonyl, 3- and 4-carboxyphenylsulfonyl-, 3-trifluoromethyl-phenylsulfonyl or 1- and 2-naphthyl among others. Thus, it seems that similarly to the matrix metalloproteinase (MMP) hydroxamate inhibitors, Clostridium histolyticum collagenase inhibitors should incorporate hydrophobic moieties at the P 1′ and P 2′ sites, whereas the α-carbon substituent may be a small and compact moiety (such as H, for the Gly derivatives reported here). Such compounds might lead to the design of collagenase inhibitor-based drugs useful as anti-cancer, anti-arthritis or anti-bacterial agents for the treatment of corneal keratitis.
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Protease Inhibitors: Synthesis of Clostridium histolyticum Collagenase Inhibitors Incorporating Sulfonyl‐L‐alanine Hydroxamate Moieties.
ChemInform, 2010Co-Authors: Andrea Scozzafava, Claudiu T. SupuranAbstract:A series of hydroxamates was obtained by the reaction of N-(4-nitrobenzyl)-L-alanine with alkyl/arylsulfonyl halides, followed by conversion of the COOH group into CONHOH. Structurally-related compounds were prepared similarly by using arylsulfonyl isocyanates, aryl isocyanates or arylsulfenyl halides instead of the sulfonyl halides. Many of the new compounds showed nanomolar affinity for the bacterial collagenase isolated from the pathogen Clostridium histolyticum.
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protease inhibitors part 5 alkyl arylsulfonyl and arylsulfonylureido arylureido glycine hydroxamate inhibitors of Clostridium histolyticum collagenase
ChemInform, 2000Co-Authors: Andrea Scozzafava, Claudiu T. SupuranAbstract:Abstract Reaction of alkyl/arylsulfonyl halides with glycine afforded a series of derivatives which were first N -benzylated by treatment with benzyl chloride, and then converted to the corresponding hydroxamic acids with hydroxylamine in the presence of carbodiimide derivatives. Other derivatives were obtained by reaction of N -benzyl-glycine with aryl isocyanates, arylsulfonyl isocyanates or benzoyl isothiocyanate, followed by conversion of their COOH group into the CONHOH moiety, as mentioned above. The 90 new compounds reported here were assayed as inhibitors of the Clostridium histolyticum collagenase (EC 3.4.24.3), a zinc enzyme which degrades triple helical regions of native collagen. The prepared hydroxamate derivatives were generally 100–500 times more active than the corresponding carboxylates. In the series of synthesized hydroxamates, substitution patterns leading to the best inhibitors were those involving perfluoroalkylsulfonyl- and substituted-arylsulfonyl moieties, such as pentafluorophenylsulfonyl, 3- and 4-carboxyphenylsulfonyl-, 3-trifluoromethyl-phenylsulfonyl or 1- and 2-naphthyl among others. Thus, it seems that similarly to the matrix metalloproteinase (MMP) hydroxamate inhibitors, Clostridium histolyticum collagenase inhibitors should incorporate hydrophobic moieties at the P 1′ and P 2′ sites, whereas the α-carbon substituent may be a small and compact moiety (such as H, for the Gly derivatives reported here). Such compounds might lead to the design of collagenase inhibitor-based drugs useful as anti-cancer, anti-arthritis or anti-bacterial agents for the treatment of corneal keratitis.
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Protease inhibitors: synthesis of Clostridium histolyticum collagenase inhibitors incorporating sulfonyl-L-alanine hydroxamate moieties.
Bioorganic & Medicinal Chemistry Letters, 2000Co-Authors: Andrea Scozzafava, Claudiu T. SupuranAbstract:A series of hydroxamates was obtained by the reaction of N-(4-nitrobenzyl)-l-alanine with alkyl/arylsulfonyl halides, followed by conversion of the COOH group into CONHOH. Structurally-related compounds were prepared similarly by using arylsulfonyl isocyanates, aryl isocyanates or arylsulfenyl halides instead of the sulfonyl halides. Many of the new compounds showed nanomolar affinity for the bacterial collagenase isolated from the pathogen Clostridium histolyticum.
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Protease inhibitors - part 5. Alkyl/arylsulfonyl- and arylsulfonylureido-/arylureido- glycine hydroxamate inhibitors of Clostridium histolyticum collagenase.
European journal of medicinal chemistry, 2000Co-Authors: Andrea Scozzafava, Claudiu T. SupuranAbstract:Abstract Reaction of alkyl/arylsulfonyl halides with glycine afforded a series of derivatives which were first N -benzylated by treatment with benzyl chloride, and then converted to the corresponding hydroxamic acids with hydroxylamine in the presence of carbodiimide derivatives. Other derivatives were obtained by reaction of N -benzyl-glycine with aryl isocyanates, arylsulfonyl isocyanates or benzoyl isothiocyanate, followed by conversion of their COOH group into the CONHOH moiety, as mentioned above. The 90 new compounds reported here were assayed as inhibitors of the Clostridium histolyticum collagenase (EC 3.4.24.3), a zinc enzyme which degrades triple helical regions of native collagen. The prepared hydroxamate derivatives were generally 100–500 times more active than the corresponding carboxylates. In the series of synthesized hydroxamates, substitution patterns leading to the best inhibitors were those involving perfluoroalkylsulfonyl- and substituted-arylsulfonyl moieties, such as pentafluorophenylsulfonyl, 3- and 4-carboxyphenylsulfonyl-, 3-trifluoromethyl-phenylsulfonyl or 1- and 2-naphthyl among others. Thus, it seems that similarly to the matrix metalloproteinase (MMP) hydroxamate inhibitors, Clostridium histolyticum collagenase inhibitors should incorporate hydrophobic moieties at the P 1′ and P 2′ sites, whereas the α-carbon substituent may be a small and compact moiety (such as H, for the Gly derivatives reported here). Such compounds might lead to the design of collagenase inhibitor-based drugs useful as anti-cancer, anti-arthritis or anti-bacterial agents for the treatment of corneal keratitis.
Matthew J Ziegelmann - One of the best experts on this subject based on the ideXlab platform.
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restoration of penile function and patient satisfaction with intralesional collagenase Clostridium histolyticum injection for peyronie s disease
The Journal of Urology, 2016Co-Authors: Matthew J Ziegelmann, Boyd R Viers, Kelly L Mcalvany, George C Bailey, Joshua Savage, Landon TrostAbstract:Purpose: Collagenase Clostridium histolyticum is approved for the treatment of Peyronie’s disease. To date, no post-release study to our knowledge has evaluated patient perceived outcomes and satisfaction. Therefore, we evaluated patient perceived experience with collagenase Clostridium histolyticum injection for Peyronie’s disease in a clinical practice.Materials and Methods: From March 2014 to July 2015, 69 patients underwent 1 to 4 series of collagenase Clostridium histolyticum injections for Peyronie’s disease at our institution. Objective changes in penile curvature as well as patient reported functional outcomes and patient perceived curvature improvements were evaluated.Results: By the time of analysis 31 patients (45%) had completed 4 trials, 47 (68%) completed 3 trials and 59 (86%) completed 2 trials. Patient reported improvements (percentage) in curvature increased with each series (trial 1—14%, trial 2—28%, trial 3—30% and trial 4—37%, p <0.05). Among those completing therapy 57% reported that ...
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Restoration of Penile Function and Patient Satisfaction with Intralesional Collagenase Clostridium histolyticum Injection for Peyronie's Disease.
The Journal of urology, 2015Co-Authors: Matthew J Ziegelmann, Boyd R Viers, Kelly L Mcalvany, George C Bailey, Joshua B Savage, Landon W TrostAbstract:Collagenase Clostridium histolyticum is approved for the treatment of Peyronie's disease. To date, no post-release study to our knowledge has evaluated patient perceived outcomes and satisfaction. Therefore, we evaluated patient perceived experience with collagenase Clostridium histolyticum injection for Peyronie's disease in a clinical practice. From March 2014 to July 2015, 69 patients underwent 1 to 4 series of collagenase Clostridium histolyticum injections for Peyronie's disease at our institution. Objective changes in penile curvature as well as patient reported functional outcomes and patient perceived curvature improvements were evaluated. By the time of analysis 31 patients (45%) had completed 4 trials, 47 (68%) completed 3 trials and 59 (86%) completed 2 trials. Patient reported improvements (percentage) in curvature increased with each series (trial 1-14%, trial 2-28%, trial 3-30% and trial 4-37%, p <0.05). Among those completing therapy 57% reported that collagenase Clostridium histolyticum injections negated a need for surgery and 52% reported restoration of penetration. Overall 81% of men perceived collagenase Clostridium histolyticum treatment as meaningful and 88% reported subjective improvements after 4 series of injections. Objective measures demonstrated a mean 23-degree curvature improvement (38%, p <0.0001). Seven patients (10%) experienced penile hematomas and no patients experienced tunical rupture. Collagenase Clostridium histolyticum reduced the need for surgery and restored penetration in the majority of patients completing 4 series of injections. It also significantly reduced the degree of objectively measured penile curvature. Subjective improvements in curvature increased with each series of collagenase Clostridium histolyticum injections as well and the majority of patients considered the therapy worthwhile. Copyright © 2016 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.
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Restoration of Penile Function and Patient Satisfaction with Intralesional Collagenase Clostridium histolyticum Injection for Peyronie’s Disease
The Journal of Urology, 2015Co-Authors: Matthew J Ziegelmann, Boyd R Viers, Kelly L Mcalvany, George C Bailey, Joshua Savage, Landon TrostAbstract:Purpose: Collagenase Clostridium histolyticum is approved for the treatment of Peyronie’s disease. To date, no post-release study to our knowledge has evaluated patient perceived outcomes and satisfaction. Therefore, we evaluated patient perceived experience with collagenase Clostridium histolyticum injection for Peyronie’s disease in a clinical practice.Materials and Methods: From March 2014 to July 2015, 69 patients underwent 1 to 4 series of collagenase Clostridium histolyticum injections for Peyronie’s disease at our institution. Objective changes in penile curvature as well as patient reported functional outcomes and patient perceived curvature improvements were evaluated.Results: By the time of analysis 31 patients (45%) had completed 4 trials, 47 (68%) completed 3 trials and 59 (86%) completed 2 trials. Patient reported improvements (percentage) in curvature increased with each series (trial 1—14%, trial 2—28%, trial 3—30% and trial 4—37%, p
Gayane Martirosian - One of the best experts on this subject based on the ideXlab platform.
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Lethal factor of Clostridium histolyticum kills cells by apoptosis.
FEMS immunology and medical microbiology, 2007Co-Authors: Jarosław Jóźwiak, Aldona Komar, Tomasz Grzela, Ewa Jankowska-steifer, Stanislaw Moskalewski, Gayane MartirosianAbstract:In this study, for the first time, Clostridium histolyticum lethal factor was purified by ammonium sulfate precipitation of culture broth, centrifugation through an Amicon filter device and hydrophobic interaction chromatography. The purified lethal factor was devoid of proteolytic activity. At a concentration of 150 ng mL(-1) the lethal factor killed 50% of HeLa cells within 24 h of exposure. Abrogation of actin filaments, activation of caspases, fragmentation of nuclear DNA as well as ultrastructural changes indicated that the cell death occurred by apoptosis. The apoptotic action of the lethal factor is in agreement with changes induced in animal tissues by administration of C. histolyticum culture medium.
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Inhibition of Clostridium histolyticum supernatant cytotoxic activity by protease inhibitors
Enzyme and Microbial Technology, 2006Co-Authors: Jarosław Jóźwiak, Aldona Komar, Ewa Jankowska, Gayane MartirosianAbstract:Abstract Clostridium histolyticum supernatant, thanks to intrinsic proteases, exerts cytotoxic effect on cells and tissues. In our study we compared the ability of five different protease inhibitors to block cytotoxic activity of C. histolyticum supertanant: aprotinin, leupeptin, phenylmethylsulphonyl fluoride (PMSF), l -1-chloro-3-[4-tosylamido]-7-amino-2-heptanone-HCl (TLCK) and chymostatin. Inhibitors of serine proteases nearly completely blocked cytotoxic effect, especially in lower supernatant concentrations. At the same time TLCK, known to irreversibly block clostripain obtained from C. histolyticum , blocked enzymatic activity of clostripain, but not cytotoxic effect of the supernatant. We conclude that the remaining cytotoxicity must be the effect of the activity of another factor that is contained in the supernatant and that can be isolated and tested in the presence of TLCK.
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Vacuolization of HeLa cells bya partially puri¢ed Clostridium histolyticum cytotoxin
2006Co-Authors: Ewa Jankowska-steifer, Aldona Komar, Tomasz Grzela, Jarosøaw J, Justyna Niderla, Stanisøaw Moskalewski, Gayane MartirosianAbstract:Clostridium histolyticum vacuolating cytotoxin was partially purified from culture broth using ammonium sulfate precipitation, gel filtration and hydrophobic interaction chromatography. The toxin caused vacuolization of HeLa cells visible under a light microscope after 2h and distinct after 8h. Transmission electron microscopy revealed the presence of numerous vacuoles, condensation of the mitochondrial matrix, increased cytoplasm density and increased amounts of heterochromatin. Apoptosis was not detected either by electron microscopy or by an apoptosis/necrosis discrimination assay with fluorescein-labeled annexin Vand propidium iodide, or DNA fragmentation assay. Calcium ion influx was detected by flow cytometry after labeling cells with Fluo-4 TM AM. Vacuolation of HeLa cells by C. histolyticum cytotoxin was inhibited by bafilomycin A1, suggesting involvement of H 1 -ATPase in the formation of vacuoles.
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Vacuolization of HeLa cells by a partially purified Clostridium histolyticum cytotoxin.
FEMS immunology and medical microbiology, 2006Co-Authors: Ewa Jankowska-steifer, Aldona Komar, Jarosław Jóźwiak, Tomasz Grzela, Stanislaw Moskalewski, Justyna Niderla, Maciej Łazarczyk, Grażyna Korczak–kowalska, Gayane MartirosianAbstract:Clostridium histolyticum vacuolating cytotoxin was partially purified from culture broth using ammonium sulfate precipitation, gel filtration and hydrophobic interaction chromatography. The toxin caused vacuolization of HeLa cells visible under a light microscope after 2 h and distinct after 8 h. Transmission electron microscopy revealed the presence of numerous vacuoles, condensation of the mitochondrial matrix, increased cytoplasm density and increased amounts of heterochromatin. Apoptosis was not detected either by electron microscopy or by an apoptosis/necrosis discrimination assay with fluorescein-labeled annexin V and propidium iodide, or DNA fragmentation assay. Calcium ion influx was detected by flow cytometry after labeling cells with Fluo-4™AM. Vacuolation of HeLa cells by C. histolyticum cytotoxin was inhibited by bafilomycin A1, suggesting involvement of H+-ATPase in the formation of vacuoles.