Antibody and Immunological Memory Responses. Rolf Zinkernagel University of Zurich, Switzerland

Transcription

1 Anti-Viral Antibody and Immunological Memory Responses Rolf Zinkernagel University of Zurich, Switzerland

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4 B cells react: Polymers (Polymeric selfantigens: collagen, DNA etc.) Monomers + LPS (CpG) 2 lymphoid organs, GC not: No B cells (low frequency or deleted) Monomers, no T help No follical structure (IgA!)

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6 nab natural IgM IgG Ab T indep. T dep. VSV 1/20 d2-4 d6-8 LCMV <1/2? d tgh25 1/40 d2-4 d6-10 µmt control/d µmt with 0.5 ml/d µmt with 2 ml/d pfu VSV i.v. Hengartner Ochsenbein Hangartner Zellweger

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11 Variable region diversity of VSV-IND neutralizing IgGs from primary (day 4/5), secondary (day 12), and hyperimmune responses (day 150) 1 2 Hyp. V H -families used by % of hybridomas V H Q52 V H 7183 V H J558

12 In vivo neutralization is dependent on a minimal serum Ab concentration in vitro in vivo Clone Subclass Avidity (M -1 ) Neutralizing 50% protective capacity (1µg/ml) concentration (dilution factor) (µg injected) G7G9E4 IgM 7x VI 22 IgG2a 9x VI 7 IgG1 5x G9 IgG2a 3x VI 29 IgG1 2x5x VI 40 IgG3 4x VI 41 IgG2a 2x VI 42 IgG2a 2x5x > 100 VI 30 IgG2a < 10 7 n.d. > 70

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14 Antibody response: ELISA vs neutr. 1 2 BSA BSA-DNP OVA BSA-DNP VSV-NJ IND-DNP DNP IND-DNP DNP nab: B limiting, neutr. a IND 1 1 ELISA a VSV NP a IND GP 2 1 DNP: T help limiting ELISA a DNP

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16 B Frequ IgG Affin. / Av. M >10 6

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18 LCMV-neutralizing titers (KL 25 nab, H-chain tg) T-indep. Hangartner, Hengartner

19 Adoptive transfer (d-1) of TgH(KL25) spl. cells Adoptive transfer (d-1) of 3x10 7 TgH(KL25) spl. cells Zellweger R., Hangartner L. et al.

20 Time dependence of LCMV-viraemia viraemia: long-term control or escape LCMV blood titer (log10 pfu) S1 S2 S3 S5 S6 S7 S8

21 polymeric AG: lgm affinity / avidity variability of AG pb

22 Role of T help pb Th where? IgA mucosal IgA no role (byst.) lam.propr. IgM no role (byst.) extra follicular IgG DNP limiting follicular nab 10-5 not limiting* follicular IgE ectoparasite spec. T dependent unclear hyper IgE serum T independent unclear *orig. antigenic sin!

23 Bachmann et al

24 B cells react: Polymers (Polymeric selfantigens: collagen, DNA etc.) Monomers + LPS (CpG) 2 lymphoid organs, GC not: No B cells (low frequency or deleted) Monomers, no T help No follical structure (IgA!)

25 A. Macpherson et al.

26 A. Macpherson et al.

27 B + T memory vs protection Quicker + better pb, pt dependent AG independent High Ab titer AG dependent plasmocytes <10d bystander act. not sufficient Co-evolutionary role (matern. Ab)

28 Memory versus protection

29 Antibody Memory after immunization with VSV a) neutralizing antibodies b) ELISA VSV-IND neutralizing IgG (-40 log 2 ) VSV-NP binding IgG (-10 log 3 ) days after immunization days after immunization

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31 Adoptive Transfer of Memory Cells V memory or naive adtf splenocytes naive or irradiated recipients +/- antigen antibody titers transfer irrad. 650 r. spleen - antigen spleen bone marrow antigen spleen VSV-neutral. IgG(-40log2) days after transfer VSV-NP (ELISA) IgG(-10log3) LCMV-WE IgG(-10log3)

32 Neutral. Ab natural Ab Ag Ag-spleen high affinity in germline protection: >10 8 l/mol, >1 µg/ml (T help) maternal Ab B cells not tolerant? monomer / multimer (crosslinking crosslinking) protection: antigen +T dependent

33 Manipulation of B response Proctective AG site (serotype not shared) Shared epitopes not protective Live vs protein dose epdemiology (Sabin) AG-dependence of Ab titer vs kinetics and cytopathogenicity of V

34 Institute of Experimental Immunology ETH and University of Zurich A. Althage M. Bachmann Z. Ciurea K. Fink S. Freigang L. Hangartner L. Hunziker U. Kalinke M. Kohler K. Lang Th. Leist M. Martinic D. Moskophidis A. Navarini A. Ochsenbein B. Odermatt D. Pinschewer HP. Pircher M. Recher T. Rülicke P. Seiler B. Senn U. Steinhoff R. Zellweger H. Hengartner

35 Thank you!

36 Adoptive Transfer of Spleen Cells: VSV VSV memory or naive adtf splenocytes naive or irradiated recipients +/- antigen antibody titers transfer irrad. 650 r. spleen + antigen spleen spleen antigen spleen VSV-neutral. IgG (-40log2) VSV-NP(ELISA) IgG (-10log3) days after transfer

37 Hypergammaglobulinemia Virus titer Doc CL 13 CL 13 Doc WE Arm WE naive Arm naive Hypergammaglobulinemia correlates with virus replication and virus persistence

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39 Protection / Mothers / Herds / Newborn Memory "idea": earlier + higher (AG-) (pb, pt) nab -: abortions / malformations nab +: herd immunity + transfer to offspr. physiological artificial vaccinations escape by variability HIV, TB: slow, T cell immunity AG driven immunopathology, no vaccines: antibiotics antivirals Emerging infections (virulence host resistance nab transfer)

40 B cell repertoire nab natural IgM IgG Ab T indep. T dep. VSV ind 1 / 20 d2-4 d6-8 LCMV < 1 / 2? d tgh25 1 / 40 d2-4 d6-10 L. Hangartner, R. Zellweger

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42 Leist Odermatt

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46 Specificity Memory Reactivity Protection

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