microbeПроблемы особо опасных инфекцийProblems of Particularly Dangerous Infections0370-10692658-719XRussian Research Anti-Plague Institute “Microbe”10.21055/0370-1069-2011-2(108)-53-56microbe-749Research ArticleИММУНОЛОГИЯIMMUNOLOGYПринципы количественной морфологической оценки безопасности противохолерных вакцинPrinciples of Quantitative Morphometric Assessment of Cholera Vaccines SafetyБугорковаС. А.BugorkovaS. A.microbe@san.ruБугорковаТ. В.BugorkovaT. V.noemail@neicon.ruКутыревВ. В.KutyrevV. V.noemail@neicon.ruРоссийский научно-исследовательский противочумный институт «Микроб»РоссияRussian Research Anti-Plague Institute MicrobeRussian Federation20112004201102(108)5356Copyright © Бугоркова С.А., Бугоркова Т.В., Кутырев В.В., 20112011Бугоркова С.А., Бугоркова Т.В., Кутырев В.В.Bugorkova S.A., Bugorkova T.V., Kutyrev V.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://journal.microbe.ru/jour/article/view/749

Разработан алгоритм оценки качества противохолерных вакцин, базирующийся на возможности применения методов количественного учета формализованных признаков, характеризующих патологический или адаптивный процесс в кишечнике иммунизированных лабораторных животных. В качестве формализованных признаков отобраны показатели: количество межэпителиальных лимфоцитов (МЭЛ), морфофункциональное состояние апудоцитов и бокаловидных клеток. Для взаимосвязанной оценки отдельных звеньев гомеостаза, морфометрические параметры представлены в виде ассоциаций показателей в форме индексов и коэффициентов. Установлено, что если определяемые показатели не выходят за определенные у соответствующих контролей пределы, то состояние адаптации организма и изменения, описываемые при гистологическом исследовании, рассматриваются как доброкачественные, характеризующие безвредность или эффективность испытуемых противохолерных вакцин.

Worked out is the algorithm of cholera vaccines quality based on application of the methods of quantitative registration of formalized parameters which characterize pathologic and adaptive processes in the intestine of the immunized laboratory animals. The following parameters were selected as formalized ones: quantity of interepithelial lymphocytes, morpho-functional condition of apudocytes and scyphiform cells. Morphometric parameters are presented as associations of indicators in the form of indices and coefficients in order to carry out interrelated assessment of separate parts of homeostasis. It was determined that in case the assessed parameters were in the limits identified in corresponding controls, adaptation condition of the organism and alterations described in histological assay were considered as benign and characterized safety or efficiency of tested cholera vaccines.

холерные вакциныморфометрические параметрыcholera vaccinesmorphometric parametersReferencesБугоркова С.А., Кутырев В.В., Бугоркова Т.В., Куличенко А.Н. Способ оценки качества вакцинных препаратов против холеры: Патент РФ 2301075. Опубл. 20.06.07 Бюл. № 17.Bugorkova S.A., Bugorkova T.V., Kutyrev V.V., Kulichenko A.N. [Method of assessment of quality of vaccine preparations against cholera]. RF Patent 2301075. 2007 June 20.Бугоркова С.А., Задумина С.Ю., Бугоркова Т.В., Лоцманова Е.Ю., Кравцов А.Л., Щуковская Т.Н., Кутырев В.В. Комплексный морфологический подход к оценке протективных свойств препаратов для специфической профилактики холеры. Журн. микробиол., эпидемиол. и иммунобиол. 2008; 1:31-4.Bugorkova S.A., Zadumina S.Yu., Bugorkova T.V., Lotsmanova E.Yu., Kravtsov A.L., Shchukovskaya T.N., Kutyrev V.V. [Complex morphological approach to assessment of protective properties of preparations for cholera specific prophylaxis]. Zh. Microbiol. Epidemiol. Immunobiol. 2008; 1:31–4.Витязева С.А., Старовойтова Т.П., Дубровина В.И., Медведева С.А., Грищенко Л.А., Шкаруба Т.Т. Морфологические изменения в иммунокомпетентных органах животных в динамике вакцинального процесса, вызванного живой чумной вакциной с иммуномодуляторами. Пробл. особо опасных инф. 2008; 3(97):50-3.Vityazeva S.A., Starovoitova T.P., Dubrovina V.I., Medvedeva S.A., Grischenko L.A., Shkaruba T.T. [Morphological alterations in immunocom- petent organs of animals in dynamics of vaccinal process caused by live plague vaccine with immunomodulators]. Probl. Osobo Opasn. Infek. 2008; 3(97):50–3.Голофеевский В.Ю. Введение в клиническую морфологию желудка и двенадцатиперстной кишки.СПб: ФОЛИАНТ; 2005. 109 с.Golofeevskii V.Yu. [Introduction in Clinical Morphology of Stomach and Duodenum]. SPb.: FOLIANT; 2005. 109 p.Ипатов Ю.П., Комарова Л.Г., Переслегина И.А., Шабунина Е.И.. Ключи к проблеме гастроэнтерологических заболеваний у детей. Н.Новгород; 1997. 217 с.Ipatov Yu.P., Komarova L.G., Pereslegina I.A., Shabunina E.I. [The Keys to the Problem of Gastrointestinal Diseases in Children]. N.Novgorod; 1997. 217 p.Кутырев В.В., Девдариани З.Л., Саяпина Л.В. Современное состояние научных исследований в области вакцинопрофилактики особо опасных бактериальных инфекций. Пробл. особо опасных инф. 2006; 2(92):18-24.Kutyrev V.V., Devdariani Z.L., Sayapina L.V. [Present status of the researches in the sphere of vaccine prophylaxis of particularly dangerous bac- terial infections]. Probl. Osobo Opasn. Infek. 2006; 2(92):18–24.Медуницын Н.В. Вакцинология. М.: Триада-Х; 2004. 448 с.Medunitsyn N.V. [Vaccinology]. M.: Triada-X; 2004. 448 p.Меркулов Г.А. Курс патогистологической техники. М.: Медицина; 1969. 423 с.Merkulov G.A. [Course of Pathogistologic Techniques]. M.: Meditsina; 1969. 423 p.Першин Б.Б., Гелиев А.Б., Толстов Д.В., Ковальчук Л.В. Система лимфоцитов ткани пищеварительного тракта животных и перорально индуцированная иммунная толерантность. Иммунология. 2001; 6:10-19.Pershin B.B., Geliev A.B., Tolstov D.B., Kovalchuk L.V. [Lymphocyte system of the animal digestive tract tissue and immune tolerance induced per os]. Immunologia. 2001; 6:10–19.Полевщиков А.В. Методологические аспекты современной иммунологии. Рос. биомед. журн. 2004; 3:17-20.Polevshchikov A.V. [Methodological aspects of modern immunol- ogy]. Ros. Biomed. Zh. 2004; 3:17–20.Санитарные правила «Государственные испытания и регистрация новых медицинских иммунобиологических препаратов» СП 3.3.2.561-96. М.: Информ. издат. центр Минздрава России; 1996. 127 с.[Sanitary Regulations “State Trials and Registration of New Medical Immunobiological Preparations” SR 3.3.2.561-96]. M.: Minzrav RF; 1996. 127 p.Саркисов Д.С., Перов Ю.Л. Микроскопическая техника: руководство. М.: Медицина; 1996. 478 с.Sarkisov D.S., Perov Yu.L. [Microscopy Technique: Manual]. M.: Meditsina; 1996. 478 p.Kabir S. Cholera vaccines: the current status and problems. Rev. Med. Microbiol. 2005; 16: 101-16.Kabir S. Cholera vaccines: the current status and problems. Rev. Med. Microbiol. 2005; 16: 101-16.Wright N.A. Aspects of the biology of regeneration and repair in the human gastrointestinal tract. Philos. Trans. R. Soc. Lound. B. Biol. Sci. 1998; 353(1370):925-33.Wright N.A. Aspects of the biology of regeneration and repair in the human gastrointestinal tract. Philos. Trans. R. Soc. Lound. B. Biol. Sci. 1998; 353(1370):925-33.Barrangou R., Fremaux C., Deveau H., Richards M., Boyaval P., Moineau S., Romero D.A., Horvath P. CRISPR provides acquired resistance against viruses in prokaryotes. Science. 2007; 315(5819):1709-12.Beloglazova N., Brown G., Zimmerman M. et al. A novel family of sequence-specific endoribonucleases associated with the clustered regularly interspaced short palindromic repeats. J. Biol. Chem. 2008; 283(29):20361-71.Beloglazova N., Brown G., Zimmerman M. et al. A novel family of sequence-specific endoribonucleases associated with the clustered regularly interspaced short palindromic repeats. J. Biol. Chem. 2008; 283(29):20361-71.Bolotin A., Quinquis B., Sorokin A., Ehrlich S.D. Clustered regularly interspaced short palindrome repeats (CRISPRs) have spacers of extrachromosomal origin. Microbiology. 2005; 151:2551-61.Bolotin A., Quinquis B., Sorokin A., Ehrlich S.D. Clustered regularly interspaced short palindrome repeats (CRISPRs) have spacers of extrachromosomal origin. Microbiology. 2005; 151:2551-61.Brouns S.J.J., Jore M.M., Lundgren M., Westa E.R., Slijkhuis R.J.H., Snijders A.P.L. Small CRISPR RNAs guide antiviral defense in prokaryotes. Science. 2008; 321(5891):960-4.Brouns S.J.J., Jore M.M., Lundgren M., Westa E.R., Slijkhuis R.J.H., Snijders A.P.L. Small CRISPR RNAs guide antiviral defense in prokaryotes. Science. 2008; 321(5891):960-4.Carte J., Wang R., Li H., Terns R.M., Terns M.P. Cas6 is an endoribonuclease that generates guide RNAs for invader defense in prokaryotes. Genes Dev. 2008; 22(24):3489-96.Carte J., Wang R., Li H., Terns R.M., Terns M.P. Cas6 is an endoribonuclease that generates guide RNAs for invader defense in prokaryotes. Genes Dev. 2008; 22(24):3489-96.Chakraborty S., Waise T.M.Z., Hassan F., Kabir Y., Smith M.A., Arif M. Assessement of the evolutionary origin and possibility of CRISPR-Cas (CASS) mediated RNA interference pathway in Vibrio choleraе O395. In Silico Biology. 2009; 9(4):245-54.Chakraborty S., Waise T.M.Z., Hassan F., Kabir Y., Smith M.A., Arif M. Assessement of the evolutionary origin and possibility of CRISPR-Cas (CASS) mediated RNA interference pathway in Vibrio choleraе O395. In Silico Biology. 2009; 9(4):245-54.Cui Y., Li Y., Gorge O., Platonov M. E., Yan Y., Guo Z. et al. Insight into microevolution of Yersinia pestis by clustered regularly interspaced short palindromic repeats. PLoS ONE. 2008; 3(7):e2652Cui Y., Li Y., Gorge O., Platonov M. E., Yan Y., Guo Z. et al. Insight into microevolution of Yersinia pestis by clustered regularly interspaced short palindromic repeats. PLoS ONE. 2008; 3(7):e2652Ebihara A., Yao M., Masui R., Tanaka I., Yokoyama S., Kuramitsu S. Crystal structure of hypothetical protein TTHB192 from Thermus thermophilus HB8 reveals a new protein family with an RNA recognition motif-like domain. Protein. Sci. 2006; 15:1494-9.Ebihara A., Yao M., Masui R., Tanaka I., Yokoyama S., Kuramitsu S. Crystal structure of hypothetical protein TTHB192 from Thermus thermophilus HB8 reveals a new protein family with an RNA recognition motif-like domain. Protein. Sci. 2006; 15:1494-9.Grissa I., Vergnaud G., Pourcel C. The CRISPRdb database and tools to display CRISPRs and to generate dictionaries of spacers and repeats. BMC Bioinformatics 2007; 8:172.Grissa I., Vergnaud G., Pourcel C. The CRISPRdb database and tools to display CRISPRs and to generate dictionaries of spacers and repeats. BMC Bioinformatics 2007; 8:172.Haft D.H., Selengut J., Mongodin E.F., Nelson K.E. A guild of 45 CRISPR-associated (CAS) protein families and multiple CRISPR/Cas subtypes exist in prokaryotic genomes. PLoS Comput. Biol. 2005; 1(6):e60.Haft D.H., Selengut J., Mongodin E.F., Nelson K.E. A guild of 45 CRISPR-associated (CAS) protein families and multiple CRISPR/Cas subtypes exist in prokaryotic genomes. PLoS Comput. Biol. 2005; 1(6):e60.Hale C.R., Zhao P., Olson S., Duff M.O., Graveley B.R., Wells L. et al. RNA-guided RNA cleavage by a CRISPR RNA-Cas protein complex. Cell. 2009; 139(5):945-56.Hale C.R., Zhao P., Olson S., Duff M.O., Graveley B.R., Wells L. et al. RNA-guided RNA cleavage by a CRISPR RNA-Cas protein complex. Cell. 2009; 139(5):945-56.Han D., Krauss G. Characterisation of the endonuclease SSO2001 from Sulfolobus solfataricus P2. FEBS Lett. 2009; 583:771-6.Han D., Krauss G. Characterisation of the endonuclease SSO2001 from Sulfolobus solfataricus P2. FEBS Lett. 2009; 583:771-6.Horvath P., Coute-Monvoisin A.C., Romero D.A., Boyaval P., Fremaux C., Barrangou R. Comparatve analysis of CRISPR loci in lactic acid bacteria genomes. Int. J. Food. Microbiol. 2009; 1(131):62-70.Horvath P., Coute-Monvoisin A.C., Romero D.A., Boyaval P., Fremaux C., Barrangou R. Comparatve analysis of CRISPR loci in lactic acid bacteria genomes. Int. J. Food. Microbiol. 2009; 1(131):62-70.Hsia K.C., Li C.L., Yuan H.S. Structural and functional insight into sugar-nonspecific nucleases in host defense. Curr. Opin. Struct. Biol. 2005; 15(1):126-34.Hsia K.C., Li C.L., Yuan H.S. Structural and functional insight into sugar-nonspecific nucleases in host defense. Curr. Opin. Struct. Biol. 2005; 15(1):126-34.Ishino Y., Shinagawa H., Makino K., Amemura M., Nakata A. Nucleotide sequence of the iap gene, responsible for alkaline phosphatase isozyme conversion in Escherichia coli, and identification of the gene product. J. Bacteriol. 1987; 169:5429-33.Ishino Y., Shinagawa H., Makino K., Amemura M., Nakata A. Nucleotide sequence of the iap gene, responsible for alkaline phosphatase isozyme conversion in Escherichia coli, and identification of the gene product. J. Bacteriol. 1987; 169:5429-33.Jansen R., Embden van J.D.A., Gaastra W., Schouls L.M. Identification of genes that are associated with DNA repeats in prokaryotes. Mol. Microbiol. 2002; 43(6):1565-75.Jansen R., Embden van J.D.A., Gaastra W., Schouls L.M. Identification of genes that are associated with DNA repeats in prokaryotes. Mol. Microbiol. 2002; 43(6):1565-75.Kamerbeek J., Schouls L., Kolk A., van Agterveld M., van Soolingen D., Kuijper S. et al. Simultaneous detection and strain differentiation of Mycobacterium tuberculosis for diagnosis and epidemiology. J. Clin. Microbiol. 1997; 35:907-14.Kamerbeek J., Schouls L., Kolk A., van Agterveld M., van Soolingen D., Kuijper S. et al. Simultaneous detection and strain differentiation of Mycobacterium tuberculosis for diagnosis and epidemiology. J. Clin. Microbiol. 1997; 35:907-14.Kunin V., Sorek R., Hugenholtz P. Evolutionary conservation of sequence and secondary structures in CRISPR repeats. Genome Biol. 2007; 8:R61.Kunin V., Sorek R., Hugenholtz P. Evolutionary conservation of sequence and secondary structures in CRISPR repeats. Genome Biol. 2007; 8:R61.Lillestøl R.K., Redder P., Garrett R.A., Brügger K. A putative viral defence mechanism in archaeal cells. Archaea. 2006; 2:59-72.Lillestøl R.K., Redder P., Garrett R.A., Brügger K. A putative viral defence mechanism in archaeal cells. Archaea. 2006; 2:59-72.Makarova K.S., Aravind L., Grishin N.V., Rogosin I.B., Koonin E.V. A DNA repair system specific for thermofilic Archaea and bacteria predicted by genomic context analysis. Nucl. Acid Res. 2002; 30:482-96.Makarova K.S., Aravind L., Grishin N.V., Rogosin I.B., Koonin E.V. A DNA repair system specific for thermofilic Archaea and bacteria predicted by genomic context analysis. Nucl. Acid Res. 2002; 30:482-96.Makarova K.S., Grishin N.V., Shabalina S.A., Wolf Y.I., Koonin E.V. A putative RNA-interference-based immune system in prokaryotes: computational analysis of the predicted enzymatic maschinery, functional analogies with eukaryotic RNAi, and hypothetical mechanisms of action. Biology direct. 2006; 1:7.Makarova K.S., Grishin N.V., Shabalina S.A., Wolf Y.I., Koonin E.V. A putative RNA-interference-based immune system in prokaryotes: computational analysis of the predicted enzymatic maschinery, functional analogies with eukaryotic RNAi, and hypothetical mechanisms of action. Biology direct. 2006; 1:7.Maraffini L., Sontheimer E.J. Self versus non-self discrimination during CRISPR RNA-directed immunity. Nature. 2010; 463(1):568-72.Maraffini L., Sontheimer E.J. Self versus non-self discrimination during CRISPR RNA-directed immunity. Nature. 2010; 463(1):568-72.Mojica F.J., Diez-Villasenor C., Soria E., Juez G. Biological significance of a family of regularly spaced repeats in the genomes of Archaea, Bacteria and mitochondria. Mol. Microbiol. 2000; 36:244-6.Mojica F.J., Diez-Villasenor C., Soria E., Juez G. Biological significance of a family of regularly spaced repeats in the genomes of Archaea, Bacteria and mitochondria. Mol. Microbiol. 2000; 36:244-6.Mojica F.J., Diez-Villasenor C., Garcia-Martinez J., Soria E. Intervening sequences of regularly spaced prokaryotic repeats derive from foreign genetic elements. J. Mol. Evol. 2005; 60:174-82.Mojica F.J., Diez-Villasenor C., Garcia-Martinez J., Soria E. Intervening sequences of regularly spaced prokaryotic repeats derive from foreign genetic elements. J. Mol. Evol. 2005; 60:174-82.Mojica F.J., Diez-Villasenor C., Garcia-Martines J., Almedros C. Short motif sequences determine the targets of the prokaryotic CRISPR defense system. Microbiology. 2009; 155(3):733-40.Mojica F.J., Diez-Villasenor C., Garcia-Martines J., Almedros C. Short motif sequences determine the targets of the prokaryotic CRISPR defense system. Microbiology. 2009; 155(3):733-40.Oost van der J., Jore M.M., Westa E.R., Lundgren M., Brouns S.J.J. CRISPR-based adaptive and heritable immunity in prokaryotes. Trends in Biochemical Sciences. 2009; 34(8): 401-7.Oost van der J., Jore M.M., Westa E.R., Lundgren M., Brouns S.J.J. CRISPR-based adaptive and heritable immunity in prokaryotes. Trends in Biochemical Sciences. 2009; 34(8): 401-7.Pourcel C., Salvignol G., Vergnaud G. CRISPR elements in Yersinia pestis acquire new repeats by preferential uptake of bacteriophage DNA, and provide additional tools for evolutionary studies. Microbiology. 2005; 151(3):653-63.Pourcel C., Salvignol G., Vergnaud G. CRISPR elements in Yersinia pestis acquire new repeats by preferential uptake of bacteriophage DNA, and provide additional tools for evolutionary studies. Microbiology. 2005; 151(3):653-63.Sorek R, Kunin V., Hungenholtz P. CRISPR - a widespread system that provides acquired resistance against phages in bacteria and archae. Nat. Rev. Microbiol. 2008; 3(6):181-6.Sorek R, Kunin V., Hungenholtz P. CRISPR - a widespread system that provides acquired resistance against phages in bacteria and archae. Nat. Rev. Microbiol. 2008; 3(6):181-6.Sturino J.M., Klaenhammer T.R. Engineered bacteriofage-defence systems in bioprocessing. 2006; 4:395-404.Sturino J.M., Klaenhammer T.R. Engineered bacteriofage-defence systems in bioprocessing. 2006; 4:395-404.Vergnaud G., Li Y., Gorge O., Song Y., Zhou D., Grissa I. et al. Analysis of the three Yersinia pestis CRISPR loci provides new tools for the investigation of ancient DNA. Adv. Exp. Med. Biol. 2007; 603:327-38.Vergnaud G., Li Y., Gorge O., Song Y., Zhou D., Grissa I. et al. Analysis of the three Yersinia pestis CRISPR loci provides new tools for the investigation of ancient DNA. Adv. Exp. Med. Biol. 2007; 603:327-38.Wiedenheft B., Zhou K., Jinek M., Coyle S., Ma W., Doudna J.A. Structural basis for DNase activity of a conserved protein implicated in CRISPR-mediated genome defense. Structure. 2009;17(10):904-12.Wiedenheft B., Zhou K., Jinek M., Coyle S., Ma W., Doudna J.A. Structural basis for DNase activity of a conserved protein implicated in CRISPR-mediated genome defense. Structure. 2009;17(10):904-12.

The authors declare that there are no conflicts of interest present.