· MHC Class II Antigen Variation Study in Selected Populations of Northern Parts of India THESIS SUBMITTED TO THE UNIVERSITY OF NORTH BENGAL, RAJA RAMMOHAN PUR FOR THE DEGREE OF DOCTOR OF PHILOSOPHY IN SCIENCE BY Sanjeev 1(p:mar Srivastava - -- ·- -·· ~;n: WI I~ 'ii'~~ t · ~L'ila~ J """Till~~- .. -- University of North Bengal, Sanjay Gandhi Post Graduate Dist. Darjeeling, West Bengal, Institute of Medical Sciences, India Lucknow-226014, India 2007 59!.2920'35Lj s 1:f4 m- 188547 29 AUG Z001 STOCKTAKING-201 1 ·I ' In memory ofmy foving 6rotfier wfw fejt for lieaven .... UNIVERSITY OF NORTH BENGAL P.O. orth Benga l Un iversity Di st. Dar:jeeling. West Bengal ltt: P"\RTJIE~'T Ot' ZOOLOGY India. Pin -734430 This is to cet1ify that the thesis entitled ''MHC Class-11 Antigen Variation Study in Selected Populations of Northern Parts of India'' is the ori ginal investigative study performed by Mr. Sanjeev Kumar Srivastava, M.Sc. under my guidance and preceptorship and has not been submitted for a degree or diploma of any university. He has carried out the work during the period2003-2006 and has ful fi lled the requirements of the degree of Doctor of Philosophy in Science (Zoology) of University of North Bengal. He is conversant with techniques and literature cited in the dissertation and carried out the work thoroughly. In character and d isposition Mr. Sanjeev Kumar Srivastava is fit to submit the thesis for Ph.D. degree. s~~~ ~o.R 7: t<..cL.L' ·~~ 01 Prof. Suraksha Agrawal Prof. T. K. Chaudhur rt Professor & Head. Professor Department of Medical Geneti cs Department of Zoology SGPGIMS, Luconow U ni versity of North Bengal (Co-l nvesti gator) (Principal In vestigator) Phone: (0353) :258:21 :2-t Fax : (0353) :2 58 15-t6 e-mail: Zoology nbu@red iffm ail.com Visit us at: \\\vw.nbu .ac.in + .9lc/Qwwfeigements 5lf{ tfiese years ofmy Pli!JJ program seems fiR! a fittfe moment. '4]:etufetf, stantiing sti{{. Just yesteraay as if. 'IFte fiefp, cooperation anti support of many assodatea airectfy or intiirectfy ena6fea me to compfete this ararwus tasK, :Many gave me a fiefping lianti anti it is my auty anti privikge to express my aeep sense ofappreciation for them. I wi{{ remain ever gratejuf to 1Jirector, Sanjay (jantifii Post (jraauate Institute of :Metfica{ Sciences, .£ucK!ww, for a«owing me not onfy to pursue my research goafs 6ut afso aevefop a scientific attitutfe. I tlianK._ tfie Institute aaministration for proviiing tfie necessary infrastructure for researcfi. I sincerefy wisli to express my t!ianifufness to tfie 'llice Cfiance«or anti 1Jean, 'University of NPrt:ft 'Benga' Sifiguri for sefecting me as a Pfi1J stutfent anti a«owing me compfete this gruefing tasK, I fed tfie fimitation of woras from insitfe in trying to express my feeilngs for my supervisor 1Jr. 'Tapas 'l(umar Cfiaualiuri, Professor, 1Jepartment of Zoowgy, 'University of :J{prtft '13engaf Sifiguri. 'Ifirougfiout my tenure, in aifferent pfiases I afways got tfie spontaneous fiefp from fiim. I express my gratituae anti sincere tlianifufness to fiim for successjuify compfeting my aissertotion work_ with liis inteifectuafguiiance anti constant inspiration. I fee{ fortune to accomp{is/i my research work_ untier tfie aynamic guiiance of my co supervisor 1Jr. _SuraR§Iia JJgrawa' ;J{eaa 1Jepartment of 'Meaica{ (jeneties, Sanjay (jamflii Post (jraauate Institute of 'Meaica{ Sciences, .£ucK!ww. 'Woras are inatfequate to express my lieartful gratitutfe to tier for i«uminative guiiance, constant inspiration, patience, encouragement, temperament anti {(jntiness to support me luring ups ani aowns in tfie course of tfie stuiy. I sfia{{ afways remain intie6teifor lier constant rMotfierfy toucfi. I am tlianifuC to 1Jean sir, 1Jr. Prof J. Pa£ (Jfetu£ '/Jept of Zoofo9!JJ, Prof.5I. '1G Cfwktavartg, Prof '/J.C. '/Jev, Prof .!'1.. !Muk_ftopaarzaga, '/Jr. S. '13arat aruf '/Jr. !M. '13afuufur, of Zoofo9!J '/Jepartment, 'llniversitg of 9-(prtft '13engaf, for tlieir vafua6fe SUiJ!Jestions luring my registration. I am afso tlianifuC to tfie non­ teaching stafffor tfieir {(jnti cooperation. I fai{ to gatfier appropriate woras to express my speciaC tlianR§ to my 'University CoCfegues !Monojit 1Jeo6nath 1Ja anti '13ik_asfi :Mitra 6ecause witliaut their fiefp ani cooperation this task couU not fiave 6een possi6fe. I rememfier tfie aays from my enro«ment, registration sampfe co«ection, practica{ fiefp etc. luring my stay at sfiguri wfien tfiese peopfe cooperate{ just {~ anything. I reaffy enjoyea with i tliem. I am afso t!Umiju{ to Slii/(J:a 'Bannerji, :Mayuk_ aruf 'Dr 5!narufita for tfieir vafua6fe suggestions. I wouit£ 6e failing in my luties if 1 lo not e;q;ress my tlia~ to my Jrieruf :faisa{ '!(!ian for a{[ liis concern, encouragements, guilance aruf assistance provitfetf luring my entire periotf. Jle not only fiefpetf aruf tauglit me {i~ a teaclier 6ut afso actetf fiR§ 6rotfier. 'IIiere are two persons, wliich neel a separate mention, 'll_ivefc Pratap Singli aruf Piyusli 'Iripatlii. I wouU fiR§ to tliank_ 'llivek_ Pratap Singli for liis ever smiling greetings wlierever aruf wfienever we ta~l aruf liscussetf rfuring tfiese years, for liis fove, fiefp, care aruf encouragements tliat I a{ways feft wfienever I was witli liim. I wouU appreciate Piyusfi 'Iri{patlii wlio continuousfy liefpetf me in tfie tedinicafities of tfie e:rperiments aruf wi{[ afways 6e tlianXju{ to liim for his continous menta{ support WordS are not alequate to tliank_ Piyusli for a{[ fie lias lone for me. I tliank_ Irufian Counci{ of :Melica{ 'l(esearcli, 9{f.w '!Jeflii for proviling me financia{ assistance luring tfie ·tenure of tliis lissertation. I was fucf}j to liave co«eagues wfw were afways reatfy to fiefp at times of neetf. I foruffy remember tfie way Sutflia li, Palma, :fa/iim ani Meena{ fiefpel me rfuring tfie fa6oratory work aruf anafysis. WordS are rea«y not enougli for tfie support of Sutflia 1~ I remem6er liow sfie tauglit me 'Dot 'Bfot aruf setting up PC9?., I am afso tlianXjuf to 'Dr Jlimansliu, :Manorama, Jl.tuC (jaurav, Jl.riz, 'Ta6rez, Jl.61iisfief(li aruf .flJ.n/Qtr for tlieir constant support. I lo not !(now fww to e;q;ress my tlia~ to 'J[.eera lili aruf Sanjay 'Bfwiya. I wi{[ afways remember tfie support, care aruf assistance of'J[.eera lirfi. Sfie fiefpel me fi~ an eUer sister. Witliaut tfie fie{p ofSanjay 'Biiaiya tliis Pli'IJ tliesis couU not liave 6een possi6fe. Jle a{ways guirfetf me rig/it tliings witli sincerety aruf fove. I wou(tf afso fiR§ to e;rpress my tlia~ to Mislira ji aruf Sliasliank_ 6/iaiya for tlieir f(jni cooperation. I am am afso tlianXjuf to '!( C Ji for liis fiefp. I am afso very mucli tlianXjuf to 5!naruf Ji aruf '!Jua Ji for tlieir secretaria{ fiefp. 'IIiis fong journey wouU liave 6een impossi6fe witliout my friendS. Specia{ tlia~ to my 6est friendS Sarufeep, Sanjeev, 'llijay aruf :Manisli. Sarufeep afways (eru{ me liis liandS at tfie time of neel aruf fijt my morafe at tfie times of lespair. 'llijay ani Sanjeev were afways tfiere witli tfieir untfemaniing care aruf spontaneous fove aruf a{ways tool( out time in spite ofliis 6eing engrossetf in tfieir own priorities, 6otli professiona[ aruf personal 'IIie togetfierness tliat ji[(el our liearts fet us 6ear a{[ tfie pressures aruf faifures of fije witli smifes. 'IIie innocent regardS teruferetf 6y :Manisli wouit£ remain memora6fe. ii (jotf cannot 6e with everyone a{{ the times. 'IIiat's wfig fre IIUUfe parents. Jt cliiftf can never thank..fris parents enougfi. Jle owes fris very l!l(jstence to tliem. Mg efforts wif{ remain incompfete if I tfo not ex:press mg gratitutfe antf intfe6tness to mg parents wfwse 6fessings stootf 6g me at a{{ otftfs antf for their constant inspiration antf encouragements antf afwags patting at times of acliievements antf a{wags correcting me at times of neetfs. It wouUf fiave 6een impossi6fe to reacli the present tfestination withaut their support. I owe a fot to mg 6rotfter :Manisli for fris constant care antf affection. I thank. mg cousin 6rotfters antf sisters Jomg, Sonu, Monu, :Himansfiu, (juria antf Sfiafu for their fove, wfiidi afwags gave me a reason to smife. I paglieartjuf thank§ to mg wife 'l(ferti for frer fove antf concern. 'During mg ups antf tfowns sfre was the person wfw gave me the reasons to smife. It wouUf fiave 6een tfiJfo:-uft withaut frer to compfete mg task._ withaut frer constant concern antf care. 'l1lortfa are rea{{g not enougli for mg son Jtryan wfrenever I fee{ tfepressetf, I usetf to see fris smife antf that mal(? me rifresliing again Mention of mg famifg is incompfete withaut mg :father in faw antf :Mather in faw. I specia{{g thank§ to Jl.nkj.t, Pratik§fia, Sfiruti antf Smriti. I thank. tliem for their everfasting fove antf care. I am thanifuC to the (jotf Jtfmigfitg, for putting sucli wontferjuC peopfe in mg fije antf 6estowing Jlis Warmth antf Jtjjection througli tfiem. Last 6ut not feast, I sincerefg fi/(? to tfetficate this thesis to mg fate eftfer 6rother !!Qtpesfi. iii Contents Pages 1 Introduction 1-26 2 Review 27-55 3 People 56-87 4 Materials and Methods 88-117 \~ 5 Observations 118-143 6 Discussion 144-196 7 Summary 197-200 8 Bibliography 201-222 Appendices (A-1) 223-249 Publications 250 ~ .9lb6reviations ddH20 Double distilled water. DNA Deoxyribonucleic Acid. EDTA Ethylenedi-amine tetra acetic acid. EtBr Ethidium bromide. FsT Inter population inbreeding coefficient. FT Intra population inbreeding coefficient. gms Grams. GsT Coefficient of population differences. HLA Human leucocyte antigens. HPLC High pressure liquid chromatography grade. lAM Infinite allele model.