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`PFIZER EX. 1103
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`PFIZER EX. 1103
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`PFIZER EX. 1103
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`THE JOURNAL
`
`OF
`
`EXPERIMENTAL MEDICINE
`
`EDI
`
`RENE DUBOS
`HENRY
`
`TED BY
`MACLYN McCARTY
`G. KUNKEL
`
`ADVISOR
`VINCENT G. ALLFREY
`ALEXANDER G. BEARN
`I}ARUJ BENACERRAF
`1{UPERT E, BILLINGHAM
`ALBERT H. COONS
`F'RANK J. DIXON
`HARRY EAGLE
`HAROLD S, GINSBERG
`
`Y EDITORS
`JEROME GROSS
`JAMES G. HIRSCH
`FRANK L. HORSFALL, Jn.
`COUNCILMAN MORGAN
`HANS J. MULLER.EBERHARD
`CHANDLER A STETSON
`JONATHAN W. UHR
`W. BARRY WOOD, Jn.
`
`e'
`fu'kfrttr
`Univusity
`
`)>)?
`
`Vor-uup 132
`1970
`
`PUBLISHED MONTHLY BY
`THE ROCKEFELLER UNIVERSITY PRESS
`
`PFIZER EX. 1103
`Page 3
`
`

`

`Copymcsr @ 1970 sv Tm Rocrrrsr,r.en UNrvrnslrv pnrss
`
`cot
`
`No. 1,.
`
`G. L. AsnpnsoN and \[.. Zvustll.
`The role of l1'mphocytes and ma
`mechanism of their interaction.
`Benny R. Broou, Lurs JrurNrz, an<
`for enumerating antigen-sensit
`sitivity.
`Enwrw Drnurn and Mexc FnrnuaNl
`the imrnune response in vitro
`nomenon of immunological tole
`Ronnnr C. Sracrn and Joosr J. Or
`macrophages and lymphocytes
`l1'nrphocl'tes.
`D. C. BrN;eurn and W. O. Wtircrn
`unresponsiveness to bovine serr
`tive and qualitative response t,
`Bncnn ANtBnssoN. Studies on thr
`the single antibody-forming cell
`after imnrunizaLion .
`Groncro ToNrnrrl, Mrcnenr B. A. O
`effect of induced chronic viral ir
`of New Zealand mice. .
`H. \Vrczurr and O. X{ArorA. Separ;
`cells by antigen-coated colurnn
`membrane antibodies as analvz
`S. Far Cunux, Hnrlrur H. Henr.r,
`Kneusn, Pnrrn A. Tolrasuro,
`on the pathogenesis of fever. )
`pyrogen-producing capacity of
`D. G. Grurrow, G.A. Tnrrs, and (
`bodl'production with cells from
`Susen Zorr.q, Jorr. Buxnarnr, E.
`Sl'nthesis and assembly of imr
`plasmacytes. I. tr4yelomas proc
`J. Wavlin Srnru.rrr.r and R. E. Bur
`host disease in Syrian hamste
`description and studies on its I
`
`PFIZER EX. 1103
`Page 4
`
`

`

`CONTENTS
`
`No. 1, Jurv 1, 1970
`
`IFELLER UNlvenstrv [)nl:ss
`
`44
`
`66
`
`77
`
`89
`
`"oot
`
`1
`
`16
`
`31
`
`G. L. Asnsnsorq and IL ZnMser.A. Contact sensitivity in the mouse. ,U.
`'fhe role of lymphocytes and macrophages in passive transfer and the
`mechanism of their interaction.
`B:erv R. Broou, Lurs JrueNnz, and Putlrr I. Nlancus' A plaque assay
`for enumerating antigen-sensitive cells in delayed-type hypersen-
`sitivity.
`ER\vrN DENon and Mflrc FErlueNw. Antibody-mediated suppression of
`the immune response in vitro. II. A new approach to the phe-
`nomenonof immunologicaltolerance.'.'
`Rosnrr c. ssrcnn and Joosr J. OnenNronn. Synergistic interaction of
`nacrophages and lymphocytes in antigen-induced transformation of
`" "
`lymphocytes
`D. c. BnNl,rurN and w. o. wnrcrr. The termination of immunological
`unresponsiveness to bovine serum albumin in rabbits. I. Quantita-
`.
`tive and clualitative response to cross-reacting albumins '
`Brncnn ANornssor.r. Studies on the regulation of avidity at the level of
`the single antibody-forming cell. The effect of antigen dose and time
`"""
`afterimmunization.
`Groncro ToNtnttl, Mrcuant B. A. OrnsroNn, and Fn'rNr J' Drxorv' The
`effect of induced chronic viral infections on the immunologic diseases
`r-rf New Zealand' mice. .
`H. WrczErr ancl o. MirruA. Separation of normal and immune lymphoid
`cells by antigen-coated columns. Antigen-binding characteristics of
`membrane antibodies as analyzed by hapten-protein antigens..:.. 110
`S. I'er Cunux, Hnruur H. HenN, Doucres II. Nlooru, DoN'lLn N'
`Knausn, PnrBn A' Touasuro, and W. B.rnnv Wool, Jn' Studies
`on the pathogenesis of fever. XX. Suppression and regeneration of
`' 127
`pyrogen:producing capacity of exudate granulocytes'. " '
`D. G. GrLuouR, G. A. THnrs, and G. J. Tuonnncrr' Transfer of anti-
`body proch.rction with cells from bursa of Fabricius
`" " " 134
`Susex Zo[r, Jonr Buxn.lulr, E' C' Fr,rNrtrw, and M' D' Scn'lnrn'
`Sl.nthesis and assembly of immunoglobulins by malignant human
`piasmacytes. I. Nlyelomas producing.y-chains and light chains. . . 1'18
`J. lVavNr SrnerrnrN and R. E. BrrrrNcneu. An analysis of graft-versus'
`host disease in Syrian hamsters. I. The epidermolytic syndrome:
`clescription and stuclies on its procttrement'
`" " " " 163
`llI
`
`PFIZER EX. 1103
`Page 5
`
`

`

`r
`
`PAGE
`
`tv
`
`CONTENTS
`
`J. WnvNo Srnrr,BrN and R. E. BrruNcrra.u. An anall,sis of graft-versus-
`host disease in S1'rian hamsters. II. The epidermolytic syndrome:
`studies on its pathogenesis.
`....... 181
`
`No. 2, Aucusr 1, 1970
`L.L.Lvzztn, R. M. Tosr, and A. O. C.qnsoNene. Electrophoreticalll.
`homogeneous antibody' synthesized by spleen foci of irradiated
`repopulated mice. . .
`Ter TB Wu and Ervru A. Kan.q.r. An analysis of the sequences of the
`variable regions of Bence Jones proteins and myeloma light chains
`and their implications for antibody complementarity...
`Jews H. GonnoN and RrN6 Dueos. The anaerobic bacterial flora of the
`mouse cecunr.
`Devrp H. KRrz, Wrnrau E. Paur, EnMolro A. Goror, and Banu;
`BcNacBnn-q.r. Carrier function in anti-hapten immune responses.
`I. Bnhancernent of primary and secondary anti-hapten antibody
`responses by carrier preimmunization. , .
`lVrrrralr E. Peur, Davrn H. Kerz, Eouoxo A. Goror, and Blnu;
`BpwecBnn-e.n. Carrier function in anti-hapten immune responses.
`II. Specific properties of carrier cells capable of enhancing anti-
`hapten antibody responses.
`Ronrnr C. SrnnNps. Host defense against bacterial endotoxemia:
`mechanism in nornral animals
`HnNny R. Hnc.lno. Synergism of thymus and bone mal.row in the pro-
`duction of. graf,t-aersu.s-host splenomegaly in X-irradiated hosts. , . . . .
`Pnrnn C. Henpnr. Human plasma alpha 2-macrogiobulin. An inhibitor of
`. plasma kallikrein.
`Jeues C. Krnxrov, PBnny E. TnrenwBll, and EpwrN S. LrNNox.
`Antigen-specific sy'nergism in the immune response of irradiated mice
`,
`given rnarrow cells and peritoneal cavity cells or extracts.
`Devm Osone. Some physical and radiobiological properties of immu-
`nologicalll'reactive mouse spleen cells. . .
`No. 3, SBprBlrnnn l, 197(l
`L. \A/. Cr-Brt and P. A. Sruer,r,, Jn. Phylogeny of immunoglobulin structure
`and function. V. Valences and association constants of teleost anti-
`bodies to a haptenic determinant.
`RrcHenl J. M,rxcr and l\4rcHa,or, R. l\{anonvev, Jn. f'he in vitro trans-
`formation of frozen-stored l1'mphocytes in the mixed l1'mphocvte
`reaction and in culture with ph)'tohemagglutinin and specific antigens.
`l\{enrrnB Y. K. AnusrRoNG, ERNST Grrrcrilr.cNx, HnrcA GrnrcuuaNN,
`
`199
`
`2rl
`
`251
`
`261
`
`283
`
`300
`
`Jl/
`
`329
`
`353
`
`368
`
`385
`
`401
`
`LonRebTn Brr,rorrr, Jann
`Scnwanrz. Chronic alioger
`phonras.
`Nonr- L. WanNnn and LBoNont
`munity to maternalll' derive
`FnBo G. Guoar, T. N. Hennrs, S
`Studies on antibody-produc
`7S antibody-producing cells.
`Juorrn C. Knncur, Gnn,q,ro Scn
`biological properties of pneu
`its capsular polysaccharide. .
`Dov L. Bonos and KBuNnrn S. !
`granuloma formation and d
`a soluble factor isolated fron
`H. G. KuNrnr, F. G. Josrrr, G
`variants of 7G4 globulin. A
`"yG globulin. . . .
`R. J. Nonru. The
`macrophages to
`tion. . .
`R. J. Nonrn. Suppression of ce
`antimitotic drug. Further r
`press imnrunity .
`Her B. RrcnrnsoN, H,q,norn
`Cutaneous basophil hyperser
`reaction, general characteris'
`Henoro F. Dvonax, ANN X{. I
`RrcnnnsoN, Sror.,rny Lrsxor
`taneous basophil hypersensit
`description
`Osras Sruruaw, Emroxo J. Yu
`thvmus function. I. Coop
`lynphohernopoietic cells in
`nrice. . .
`Osr.rs SruruaN, EouoNn J. Yu
`thymus function. IL Coope
`hemopoietic liver cells wiih
`
`relative im
`the express
`
`No.4
`ElN.l Mozss, G.M. Sreerun, a
`genetic control of immune
`
`PFIZER EX. 1103
`Page 6
`
`

`

`CONTENTS
`
`PAGE
`
`448
`
`LorseNn Botnortr, JeNrun ANlnr-Scnwlntz, and Ronrnr S'
`Sfirwenrz. chronic allogeneic drsease. II. Development of lym-
`' " 117
`phomas'
`^ronl t. w.,lnNon and LnoNonr A. HERzENBERG. Tolerance and im-
`nrunity to maternally derived incompatible IgGz"-globulin in mice. . 440
`IhBo G. ()unar, T. N. H.qnms, susANNA Hltnts, and Kr-,A.uS HrruuBrnn.
`Stuclies on antibody-producing cells. I. ultrastructure of 19S and
`TS antibody-producing cells '
`luorrH C. KNrcnt, Gnnrrrp ScmrlueN, and Ronnnr Ausrnr'q'N' Some
`biological properties of pneumococcus Type 37 and the chemistry of
`" 475
`it..u!r.,1*.polysaccharide""
`DovL.nonosu,'dK"**u'us.WenneN.Delayedhypersensitivity-type
`gr:rnuloma formation and dermal reaction induced and elicited by
`. . 488
`i soluble factor isolatecl from Schistosoma mansoni eggs .
`H. G. KuNxBl, F. G' Josuw, G. M' PaNr'r, and J' B' Nervrc' Genetic
`variants of "yG4 globulin. A unique relationship to other classes of
`lG globulin.
`n. ;. Nonru. The relative importance of blood monocytes and fixed
`macrophages to the expression of cell-mediated immunity to infec-
`....52t
`tion...
`R. J. Nonrn. Suppression of cell-mediated immunity to infection by an
`antimitotic drug. Further evidence that migrant macrophages ex-
`""
`pressimmunitY. .
`H.rr B. RtcHetsoN, Hltorl F. Dvoner, and SmNnv Lnsrowtrz'
`cutaneous basophil hypersensitivity. I. A new look at the Jones-Mote
`reaction, general characteristics. . . '
`H'rnorl F. Dvontr, ANN M. Dvonlr, Br.tNcrrn' A. SrarnsoN, H.qr B.
`Rtcrrnnsow, SroNrv Lnsxowtrz, and Monnrs J' Kanrovsxv' Cu-
`taneous basophil hypersensitivity. II. A light and electron microscopic
`description.
`Osres SruiuAN, Eoltoxo J. YuNrs, and Roncnr A' Gooo' Studies on
`thymus function. I. Cooperative efiect of thlmic function and
`lymphohemopoietic cells in restoration of neonatally thymectomized
`mlce..
`Osras Sruruan, EurtoNn J. Yunrs, and Rosnnn A' Gooo' Studies on
`thymus function. II' Cooperative effect of newborn and embryonic
`hemopoietic liver cells with thymus function
`No. 4, Ocrosnn l,1970
`EDNA MozEs, G. IVI. SrnAnrn, and NftCnirnl SELA. cellular basis of the
`genetic iontrol of immune responses to synthetic polypeptides. I.
`
`508
`
`535
`
`5.r6
`
`558
`
`583
`
`601
`
`; r
`
`. An analvsis ol graft-aersus'
`The epidermolrtl srndlome:
`
`t, l97o
`RBoNARA. ElectroPhoreticallY
`by spleen foci of irradiabed
`
`181
`
`199
`
`2ll
`
`nal-vsis of the seqr-rences of the
`:ins and myeloma light chains
`conrplementaritY. . .
`: anaerobic bacterial flora of the
`. 251
`DMoND A. (ioror, and Benul
`lnti-hzrpten immune resPonses'
`condary anti-haPten antibodY
`."
`)n. ...
`DlroND A. Goror,, and Beru;
`inti-hapten immune resPonses.
`lls capable of enhancing anli-
`. . ..283
`Lgainst bacterial endotoxemia:
`
`261
`
`300
`
`us and bone marrow in the Pro-
`galy in X-irradiated hosts . 3I7
`2-macroglobulin. An inhibitor of
`
`329
`
`,vErL, and Epwrlr S. LPNNox'
`nune response of irradiated mice
`' 353
`:avity cells or extracts
`obiological properties of immu-
`. J68
`.
`r.s.....
`
`:nn 1, 1970
`;eny of immunoglobulin structure
`:iation constants of teleost anti-
`
`385
`
`RDTNEY, JR. The in vitro trans-
`cytes in the mixed lYmPhocYte
`nagglutinin and specifrc antigens. '401
`ETCHMANN, Hnrcl Gr-ntcnulNwt
`
`PFIZER EX. 1103
`Page 7
`
`

`

`vl
`
`CONTENTS
`
`CONl
`
`Differences in frequency of splenic precursor cells specific for a s1.n-
`thetic pollpeptide derived from multichain pollproline ([T, G]-
`. Pro--L) in high and low responder inbred mouse strains
`G. Cuoxowrcz, G. M. Snrenrin, and T. Iro. Cellular difierentiation
`of the immune system of mice. VI. Strain differences in class dif-
`ferentiation and other properties of marrow cells. . .
`J. P. DrrnEv, Naxcv L. Mnrnn, and J. K. FnrNxrL. The Toxoplasma
`gondi.i oocyst from cat feces. .
`SrppnrN L MonsB and Bnucl A. BennoN. Studies on the leukocytosis
`and l1'mphocytosis induce d by Bord,etella pertus sts. III. The distribu-
`tion of transfused lvmphoc)'tes in pertussis-treated and normal
`mice..
`Fn.q.Nr R. ScHum, IvaN I\{. Ronr, and Manra J. Rocrn. Complement
`' fixation by a two-componenl antibody systern: immunoglobulin
`G and immunoglobulin M anti-globulin (rheumatoid factor). Para-
`doxical efiect related to immunoglobulin G concentration
`Geny D. Ovrnrrrx.l and Enw,rnl A. Monrrurn, Jn. Studies on the
`relationship between the production of bacteriocines by Group A
`streptococci and acute glomerulonephritis. .
`Cnrso BreNco, Rrcn.q.no PATnrcr, and Vrcron NusseuzwErc. A popula-
`tion of lymphocytes bearing a rnembrane receptor for antigen-
`antibody-complement complexes. I. Separation and characteriza-
`' . tion
`AreN E. Bnan and R. E. BmrNcnau. Implantation, transplantation,
`and epithelial-mesenchymal relationships in the rat uterus.
`Mrcnenr Porrnn and Rosr LrnsoRMaN. Common individual antigenic
`determinants in five of eight BALB/c IgA mt'eloma proteins that
`bind phosphoryl choline.
`Gurro Brozzr, RrcueRD Asorsrv, Rosn LrasBnMAw, CTAUoE SrrFl-EL,
`DnNrsB Mourox, and B.l.nu1 BnNeconna.r. Serum concentrations
`and allotlpes of immunoglobulins in two lines of mice geneticalll
`selected for "high" or "low" antibody synthesis.
`B. SneNNox DAwes, J. E. Scorr, and ArBxeNnrn Ci. BnenN. Further
`sludies on metachromasia in cultured human fibroblasts. Staining
`of glycosaminogll,6ans (mucopoll'saccharides) br. Alcian blue in salt
`solutions.
`Nrrr, R. CoopBn and.','Eaws J. I\{rinon-EsnnH,{np. The reaction
`mechanism of hunran C5 in imnrune hemolysis.
`Rarpn vAN FuRTrr, J,lrr.ms G. Hrnscn, and n4nnrna. E. Fnnonxo. N{or-
`phologl' and peroxidase cytochemistrl' of mouse prolronocytes,
`monoc\-tes, and n'racrophages. . .
`
`PAGE
`
`613
`
`623
`
`636
`
`663
`
`673
`
`691
`
`702
`
`721
`
`lJt
`
`765
`
`I lJ
`
`791
`
`Rerpn vAN FuRTH and l\'Ie.nrrN.a, i\{.
`kinetics of promonocytes and mor
`D. Snr-r-rN, l\'I. Kcivnnv, U. Rornon,
`plemer.rt fixation b1' c1'totoxic an
`
`No.5, Novr
`V. S. Bvsns and E. E. SEncaB.z. I
`Parall'si5 in vitro '
`M. VaN \\/rNrue and L. LrvY. Fur
`serum sickness cholesterol-inducec
`PBrpn B. LeMsu{r and Howenn A. F
`ponents of the allograft reactic
`allografts.
`Narn.q,N Tn.qnrN and MYn'q. SulLL.
`properties of a thYmus humorl
`petence on lrmphoid cells. .... . .
`Orro Gorzr and HeHs J. X{iiurn-.
`b1'complement in the absence of a
`Pnrrn J. McCunacu. The abrogat
`in rats b1'uteans of the transfer ol
`Wrsrrv W. Burlocr and N{envrN I
`secondary anti-hapten response.
`antigen
`N. K. B. Dev, H. Gnwunz, R. Jo
`Goon. Cornplement and comPl
`tebrates and invertebrates... . . . .
`Bnttco W. Bnraxr and Ar,rnno Nrsox<
`idiotlpic antibodies. IV' Inhibitio
`of anti-hapten antibody with its a
`H. F. Jrr;oEBr{oY. The effect of ht
`on lymphocytes of thymus and n
`KNur D. KNuosBlr, LBwrs K. DaHl.
`Mlnrnl Hrrxo. and Gponcn Ll
`ingestion. Inheritance of h1'perten
`BENt'rsuro Pnnxrs, Lucr,qua Fomi
`globulin spots on the surface of rt
`VorxE,n Scurnnrrecunn and Kr,.r,us I
`body class in a s1's1sm of cooPera
`R. V. Br.eNprtq. Mechanisms of reco
`tion: nousepox. L The effects of
`
`PFIZER EX. 1103
`Page 8
`
`

`

`i
`
`CONTENTS
`
`vii
`
`PAGE
`
`813
`
`829
`
`rcursor cells speci6c for a sYn-
`Itichain polyproline ([T, G]-
`. 613
`inbred mouse strains. .
`Iro. Cellular differentiation
`itrain differences in class dif-
`narrow cells. . .
`-. FR.eNror. The ToroPlosmo
`
`623
`
`636
`
`663
`
`6i3
`
`69+
`
`702
`
`721
`
`r. Studies on the leukocYtosis
`lla pertussts.III. The distribu-
`pertussis-treated and normarl
`
`lenn J. Rocxrt. ComPlement
`rody system: immunoglobulin
`.lin (rheumatoid factor). Para-
`Iin G concentration.
`oRTTMER, JR. Studies on the
`of bacteriocines by GrouP A
`ritis
`:ron NussBNzwEIG. A PoPula-
`nrbrane receptor for antigen-
`Separation and characteriza-
`
`lrnplantation, transPlantation,
`hips in the rat uterus.
`Common individual antigenic
`/c IgA my'eloma Proteins that
`
`LteenRltln, Creuur Strrret,
`cERR.\F. Serttm concentrations
`n two lines of rnice geneticalll'
`ly synthesis
`LEXANDER G. Br.lRN. Fttrther
`'ed human fibroblasts. Staining
`charides) by Alcian blue in salt
`
`r.ren-EsnnHlro. The reaction
`e hemolysis
`rd llnnrn,r E. Fnoonxo. IIor-
`,istry of motise Promonocytes''
`
`i \')
`
`765
`
`I lJ
`
`791
`
`Rarpn vAN FURTH and iV.rnrrN'r' I\I' C' DEsserHorr-DEN DULK'
`kinetics of promonocl'tes and monocytes in the bone marrow'
`D. SenrN, NI. Kov,rnv, U. Rorurn, and K' Rorsen' Intrarenal
`plement lixation by cytotoxic antibodies
`
`The
`
`com-
`
`Slrer.r. Studies on some physicochemical
`humoral factoi conferring immunocom-
`
`Lysis of erythrocytes
`
`, uu,*, ancr E. " itJ;"lJ-1"ffi,1;tlll ..u.,,u, or immune
`".
`paralvsis in vitro.
`,\1. iAx ivt^*tu and L. LBvy. Further studies on the reversibility of
`serun.r sickness cholesterol-induced atherosclerosis''
`prrnR B. l,Arasenr and HOw,lr.p A. FnaNx. Cellular and vascular com-
`ponents of the nllograft reaction. Evidence from returned skin
`lullografts. . .
`NrrH.rN 'fnlrNtrg and MYn"\
`properties of a thYmus
`petence on lymPhoid cells.
`Orro Gorzr and HeNs J' lIUr'mn-EBERt{ARD'
`b1, complement in the absence of antibody '
`rarrn;.NlcC]urr.c'cH.Theabrogationofsheeperythrocytetolerance
`in rats b1' means of the transfer of allogeneic lymphocytes" "
`wl:sr-cy w. Bu[ocr and M,rnvrls B. RrrreNsrnc. In vitro-initiated
`seconclary antlhapten response. II. Increasing cell avidity for
`iLntigen
`N. K. B. D,rv, H' Gewunz' R. JoneNNsEN, J' FlNstlo, and R' A'
`6ooo'Complementandcomplement-Iikeactivityinlowerver-
`tebrates and invertebrates..
`Bnuc.E w. BnlExr and Arrnno NrsoNonr. Quantitative investigations of
`idiotypic antibodies. IV. Inhibition by specific haptens of the reaction
`of anti-hapten antibody with its anti-idiotypic antibody
`H. F'. Jnn;rrnnov. The effect of heterologous anti-lymphocyte serum
`on l-v-pho.ytes of thymus and marrow origin
`Kxuo D. KNulsnN, Lnwrs K. Denr,, KrITn T)toursou, JrNcnr Iwer'
`I[.c.nrH.{ HErNn, and GBORCT Lnru. Effects of chronic excess salt
`ingestion. Inheritance of hypertension in the rat
`'
`Br^-vlr'quro PnxNrs, LucrtNr FonNt, and Lutsa Alr'txrn' fmmuno-
`globulin spots on the surface of rabbit lymphocytes'
`vorrnn Scnrnnuecuen and Kr.lus RAJEWSKY. Determination of anti-
`bod-r' class in a system of cooperating antigenic determinants
`R. V. BieNorx. llechanisms of recovery from a generalized viral infec-
`tion: monsepox. I' The effects of anti-thymocyte serum'
`" ' 1035
`
`8-t5
`
`858
`
`868
`
`885
`
`898
`
`916
`
`926
`
`94t
`
`95l
`
`963
`
`976
`
`1001
`
`l0l9
`
`PFIZER EX. 1103
`Page 9
`
`

`

`VIII
`
`CONTENTS
`
`c{
`
`PAGE
`
`1055
`
`1071
`
`No. 6, Drcr,unnn 1, 1970
`Inux R. ComN, Lenv Sravv, and Mrcn.ln Frtoul,lv' Glucocorti-
`coids and cellular imrnunity in vitro. Facilitation of the sensitization
`phase and inhibition of the efiector phase of a l1'mp[ec1'te anti-
`fibroblast reaction.
`EnnN BonnNrnnuND, YuJr Hoxna, Mrrnnno StotNctLss, and A,qnou
`BsNnrcE. Studies of DNA-induced heritable alteration of maur-
`malian cells. .
`Josrur D. Scnur,M,lx and KerrnvN H. Bnlnmv. The metabolism of
`amino acids, peptides, and disulfides in lysosomes of fibroblasts cul-
`. . . . 1090
`tured from normal individuals and those with cystinosis.
`Mrcntnr- IlrpnlsBncnn and Monrv E. Sroorr. Predicted and un-
`predicted cross-reactions of an acetylphosphogalactan of Sporobolo'
`. "'
`ffiycesyeast.Il.....
`D. G. Cor,rev, A. Y. Smu Wu, and B. H. WlrsueN. Cellular difierenti-
`ation in the thymus. III. Surface properties of rat thpnus and Iymph
`node cells separated on density gradients
`Henorp C. Mrr,r,en and Gusravo Cunrowcz. Antigen-specific cells in
`mouse bone marrow. I' Lasting effects of priming on immunocl'te
`production by transferred marrow.
`Jonr G. Blsnuau, A. M. PepprNIrErMER, Jr., D.M. Grr,r,, and ANNarpr.
`. .
`A. Henrrn. Action of diphtheria toxin in the guinea pig
`M. A. VnNx.qrAcIrALAM, X{. J' KenxovsKY, H' D. Farrur, and R' S'
`Cornerv. An ultrastructural study of glomerular permeability using
`catalase and peroxidase as tracer proteins.
`I{. A. VeNrerlcneuu, R. S. CornlN, and M. J. KenNovsrv' An
`ultrastructural study of glomerular permeability in aminonucleoside
`nephrosis using catalase as a tracer protein.
`1'. Tlxeresur, E. A. Canswntl, and G. J. Tlronrncrn' Surface anti-
`gens of immunocompetent cells. I. Efiect of d and PC.1 alloantisera
`on the abilitl' of spleen cells to transfer iurmune responses.
`N. AsReltsoN, A. F. LoBucrro, J. H' JeNor, and R. S. CorneN. The
`interaction between human monocl'tes and red cells. Binding char-
`acteristics.
`N. AsnAMsoN, E. W. Grr,rexo, J. H. JeNnr, and F'. S. Rosrx. The in-
`teraction between human monoc)'tes and red cells. Specificitl- for lgG
`subclasses and IgG fragrnents'
`J. J. T. Ovnx and M. C. R-q'rn' Studies on the difierentiation of thyrrus-
`derived l1'mPhocytes
`Crrnra S. W.q.rrnns and Haxs Wrcznrr' I)emonstration of heavl' and
`light chain antigenic determinants on the cell-bound receptor for
`
`antigen. Similarities between
`antibodies produced by the sanr
`Cuenrns HuGcINs, Sor Wrsnlreu,
`of fibroblasts by allogeneic and
`lized tooth and bone.
`Wrlrreu Jonn MenuN, LEoNARD
`BoNecsRRAr. Histocompatibil
`ness in random bred Hartley str
`Kr.lus-Urnrcu HlnrunNN. Induct
`Interaction of cells of bone rnar
`Jn.l,w-Crauon BYSrnvu, Manrrx \
`Regulation of antibodl' formati
`of specific antibody bv means o.
`
`L
`
`1105
`
`1107
`
`tl22
`
`1 138
`
`1 153
`
`1168
`
`1 181
`
`tr9l
`
`t207
`
`tzr6
`
`PFIZER EX. 1103
`Page 10
`
`

`

`CONTENTS
`
`ix
`
`PAGE
`
`humoral
`
`r233
`
`antigen. Similarities between membrane-attached and
`antibodies produced by the same cell . . . . .
`Rroor. Transformation
`Cnrnrrs HucerNS, Sor Wrsslr.qN, and A' H.
`transplants of deminera-
`of fibroblasts by allogeneic and xenogeneic
`... ..1250
`lizedtoothandbone
`\\Irnun JouN lt.rnrrN, LEoNARD EttM,tN, Ine GnooN, and Banu;
`Bs,rA.crnzur. Histocompatibility type and immune responsive-
`. t259
`ness in random bred Hartley strain guinea pigs. . .
`Ki..r.us-Urnrcu H.rnruexN. Induction of a hemolysin response in vitro'
`Interaction of cells of bone narrow origin and thymic origin' ' .... 1267
`J.o.rx-Cir,ruon BvsrnYN, NIrnrrN W. Gnlr', and Joxtrtr.rx W. Unt.
`Regtrlation of antibody formation b1' serum antibody. II' Removal
`. .
`of specific antibody by rneans of exchange transfusion.
`
`1279
`
`n 1, 1970
`tlu Fnrnu,tN. Glucocorti-
`acilitation of the sensitization
`phase of a lymPhocYte anti-
`
`)RED SrErNcl,,tss, and A-AtoN
`heritable alteration of mam-
`
`IR.qDLEY. The metabolism of
`n Iysosomes of fibroblasts cul-
`,e with cystinosis. . .
`Sroprr. Predicted and ttn-
`phosphogalac tan oI S P o robolo'
`
`105s
`
`1071
`
`1090
`
`1105
`
`ll07
`
`V,rrsM..LN. Cellular differenti-
`:rties of rat thYmus and lYmPh
`nts. .
`Antigen-specific cells in
`NICZ.
`rts of priming on immunocYte
`" "' tl22
`Jn., D.M. Grr,r,, and ANN.rsar
`. ' 1138
`rin i.t the guinea Pig .
`,rv, H. D. Fenrm, and R' S'
`i glomerular permeability using
`..
`" 1153
`;eins..
`J. KenNovsrv. An
`and M.
`ty in aminonucleoside
`ermeabili
`rotein. . .
`J. Tnonnncro. Surface anti-
`ffect of 0 and PC.l alloantisera
`' 1181
`:r immune responses.
`[NDL, &nd R. S. Corn-lN' The
`-es and red cells. Binding char-
`
`1 168
`
`tor., and F. S. RosnN. The in-
`and recl cells' SPecificitY for IgG
`
`1 191
`
`1207
`
`rn the differentiation of thYmus-
`. .. .. - 1216
`. Demonstration of heav;' and
`on the cell-bound recePtor for
`
`PFIZER EX. 1103
`Page 11
`
`

`

`THE JOURNAL OF EXPERIMENTAL }IEDICINE
`;ulv t, 1970
`Vor.ulm 132. Nuunrn 1
`
`PFIZER EX. 1103
`Page 12
`
`

`

`QUANTITATIVE INVESTIGATIONS OF IDIOTYPIC ANTIBODIES
`
`IV. INHIBITION BY SPECIFIC H APTENS OF THE REACTION OF A NTI-HAPTEN
`A NTIBODY WITH ITS ANTI-lnIOTYPIC A NTIBODY*
`
`BY BRUC E W. BRIENT,t M.D. , AND ALFRED NISONOFF, PH.D.
`(From //,c Departments of Surgery and Biological Chemistry, University of Illinois at
`tl,e Medical Center, Chicago, I llinois 60680)
`
`(Received fo r publication 6 July 1970)
`
`Myeloma proteins (1) and antibodies from individual animals (2, 3) have
`been shown to possess individually specific or " idiotypic" ( 4) antigenic de(cid:173)
`terminants. For example, anti-salmonella antibodies from an individual rabbit
`have determinants that are not detec table in other immunoglobulins of that
`rabbit nor, ordinarily, in antibodies of the same specificity from other rabbits
`(3). Quantitative studies have shown that various fractions (up to 80 %) of
`purified anti-benzoate antibodies, designated D (5), from a given donor rabbit
`are reactive with anti-idiotypic (anti-D) antisera (6, 7). Precipitin lines in the
`Ouchterlony test were obtained when as little as 2% of the donor (D) popula(cid:173)
`tion was precipitable.
`Our present investigations indicate that the reactions of anti-p-azobenzoate
`antibodies (D) with their anti-idio typic antisera are strongly inhibited by
`benzoate derivatives, i.e. by specific haptens, whereas the reactions of D with
`anti-Fab antibodies are affected to a much smaller extent. The possible relation(cid:173)
`ship of icliotypic determinants to the antigen-combining site of a donor antibody
`is discussed.
`
`Materials and Methods
`
`The following methods and materials have been described previously (6, 7): preparation
`and specific purification of anti-p-azobenzoate antibodies of the IgG class; polymerization
`of these antibodies with glutaraldehyde for the purpose of immunization; preparation and
`labeling with 125IC1 of F(ab'h fragments of purified anti-benzoate antibodies (each such
`preparation containing a 20-fold excess of nonspecific,'F[ab'h fragments); preparation of goat
`antiserum specific for rabbit fragment Fe; and determination of allotypes of rabbit sera.
`Anti-allotype antisera were the generous gift of Dr. Sheldon Dray.
`
`* This wo rk was supported by grants from the National Science Foundation (GB-5424)
`and the National Institutes of Health (A l -06281 ). It was presented in part at the annual
`meeting of the Federated Societies for Experime11tal Biology and Medicine,-ApriT, 1970
`(Fed . Proc . 29:773).
`t Supported by a National Institute of Medical Sciences Surgery Academic Training Grant,
`U. S. Public Health Service grant o. GM 1930-0I A 1.
`
`951
`
`PFIZER EX. 1103
`Page 13
`
`

`

`952
`
`QUANTITATIVE INVESTIGATIONS OF ID10TYPIC ANTIBODIES. IV
`
`Percentages of 1251-F(ab'h fragments of D antibodies precipitable were determined by
`an indirect method utilizing excess goat antibody directed to the fc fragment of rabbit lgG
`to precipitate complexes of 125I-F(ab'h fragments with anti-D antibodies. The method used
`corresponds to that reported previously (7) with two exceptions. First, ovalbumin (SO µ g
`per test). was -used in place of bovine seruin albumin to minimize adherence of the labeled
`protein to glass. Serum albumin was not used because it is known to bind various small
`molecules which were tested as inhibitors. Second, controls were run by utilizing anti-dinitro(cid:173)
`phenyl (anti-DNP) antiserum in place of anti-D. (Formerly anti-ovalbumin was used as a con(cid:173)
`trol). Percentages of radioactivity precipitated in control experiments did not exceed 2.8%.
`The control value was subtracted in each series from percentage values obtained with anti-D
`antiserum .
`When small molecules such as haptens \Vere tested as competitors of the reaction of F(ab')2
`fragments of D with anti-D, the competitor was first incubated with the labeled F(ab'h
`fragments and 50 µg of ovalbumin for 2 hr at 37°C, pH 8. Anti-D antiserum was added , in(cid:173)
`cubation was continued for 1 hr, and goat anti-rabbit- Fe was then added to the mixture .
`After standing for 3 days in the refrigerator, the percentage of radioactivity precipitated was
`determined with a gamma scintillation counter (7). A minimum of 3000 counts was recorded
`for each precipitate-supernatant pair. Experiments were carried out in triplicate. All hap tens
`and other small molecules tested as competitors were crystallized from water or from an
`ethanol-water mixture, with the exception of p-nitrobenzene sulfonic acid, and used as the
`sodium salts. Solutions were adjusted to pH 8.
`Preparation of Anli-Idiotypic (Anti-D ) Antibodies.- Specifically purified anti-benzoate
`antibodies (D) from individual rabbits were injected into recipients of allotype matched to
`the donor with respect to the following specificities: al , a2, a3 , b4, bS, b6, b9, c7 , c21.
`Rabbit 9Q was immunized with purified D antibody from rabbit A25, according to the
`second protocol given in reference 6; 3- 5 mg of monomeric D were used for the initial inocu(cid:173)
`lations with Freund 's adjuvant, and polymerized D was used for subsequent intravenous
`inoculations. A pool of antiserum from rabbit 9Q was made from several bleedings which
`showed strong antibody activity by the method of indirect precipitation.
`Purified D antibodies from rabbits A4 and 1-14 were injected exclusively in the poly(cid:173)
`merized form into recipients 14E and lOX, respectively, and D antibody from rabbit AS was
`injected into 3 recipients, 7 A, 7C, and 7D. Two subcutaneous inoculations of 3 mg each in
`complete Freund's adjuvant were given 3 wk apart. These were followed by intravenous in (cid:173)
`oculations at intervals of 2-4 wk. Bleedings were taken 5-8 days after an injection . Antisera
`from individual recipients which gave strong reactions by the method of indirect precipitation
`were pooled. Evidence that antibodies produced were directed to idiotypic determinants, and
`not to allotypic or hidden determinants, has been described in detail for D antibodies of rab(cid:173)
`bits AS and A25 (7, 8). This evidence includes the failure of sera taken from donor rabbits
`prior to immunization to inhibit reactions of anti-D with 125I-F(ab')2 fragments of D, the loss
`of inhibitory capacity of whole D serum after precipitation of antibenzoate antibodies, and
`the specificity of the reactions when various anti-D sera and D antibodies were utilized.
`Similar results were obtained in a large number of preliminary control experimen ts with
`the D antibodies of rabbits I-14 and A4.
`The percentages of F(ab'h fragments of specifically purified anti-p-azobenzoate anti(cid:173)
`bodies that were precipitable by anti-D antisera, using the indirect method, were: rabbit AZ5,
`57 ±3% ; rabbit AS, 37 ±2% ; rabbit l - 14, 31 ±1 % ; rabbit A4, 23 ± 1% .
`
`The effects of specific haptens and other small molecules on the reactions of
`125I-F(ab'h fragments of D antibodies with anti-D an lisera are shown in Tables
`
`RESULTS
`
`PFIZER EX. 1103
`Page 14
`
`

`

`BRUCE W. BRIENT AND ALFRED NISONOFF
`
`953
`
`TABLE I
`Rjfect of 1-foplens and Other S11wll Molec11les on the Reaction of 120I-F (ab')2 Derived from D
`A nlibodies of Rabbit A Z5 with Anti-D Sernm*
`
`Compet it or
`
`Final molar concentration of competi tort
`
`t.6 X 10- ,
`
`5 X 10-•
`
`5 X 10- •
`
`Rel. K (9)
`
`p-(p' -hydroxy )-phenylazoben-
`zoatc
`benzoate
`p-nitrobenzoate
`m-nitrobenzoate
`o-nitrobenzoate
`p-am inobenzoate
`111-aminobenzoate
`o-aminobenzoate
`p-bromobenzoate
`111-bromobenzoate
`o-bromobenzoate
`p-chl orobenzoate
`m-chlorobenzoate
`o-chlorobenzoate
`p-iodobenzoate
`o-iodobenzoate
`p-methylbenzoate
`m-methylbenzoate
`o-methylbenzoate
`
`Sodium acetate
`Potassium iodide
`Potassium bromide
`p-(p' -dimethy lamino)-phenyl-
`azobenzene sulfonate
`
`" ' l -F (ab' ), pru ipitaled, % of co11lrol§
`57 (! )
`6 1 (5)
`
`74 (4)
`
`73 (4)
`69 (2 )
`74 (3 )
`90 (1)
`88 ( I )
`93 (3 )
`76 (1)
`75 (1)
`8 1 (2)
`95 (1)
`64 (2 )
`78 (7 )
`8 1 (3)
`64 (2 )
`87 (I)
`71 (2)
`87 (2 )
`92 (2 )
`
`94 (I)
`98 ( I )
`97 (1)
`112 (1)
`
`90 (4 )
`70 (3 )
`86 (3)
`98 (1)
`96 ( \ )
`91 (! )
`85 (l)
`86 ( I )
`98 (\)
`103 (1)
`77
`(2 )
`83 (3)
`97 (1)
`75 (2 )
`97 (1)
`81 (I)
`97 (3)
`100 (1)
`
`99 (1)
`102 (2 )
`102 (1)
`104 (1)
`
`95 (1)
`82 (2)
`85 (4)
`89 (2)
`97 (2 )
`96 (3)
`93 (2)
`78 (2)
`93 (4)
`96 (1)
`(2)
`91
`89 (2 )
`87 (5)
`90 (1)
`
`91 (I)
`102 (2)
`102 (I)
`
`99 (t)
`101 (3)
`95 (1)
`94 (2)
`
`22
`
`1.0
`11 .5
`0.4
`<0 .1
`0.9
`0.3
`1. 5
`5 .0
`1.3
`0.1
`3.7
`0.8
`0.2
`
`2 .6
`0 .7
`0.1
`
`• Anti-D serum was from rabbit 9Q. The indirect method of precipitation was used.
`Each test contained 0.5 µg 125! -F(ab'h of the D antibody, 9.5 µg nonspecific F(ab'l2, and SO
`µg ovalbumin in a final volume of 0.31 ml. lO µl of anti-D serum and 0.4 ml goat anti-rabbit
`Fe were then added. In the absence of competitors 57 ± 3% of the radioactivity was pre(cid:173)
`cipitated wi th anti-D serum and 1.2% in the control utilizing lO µl anti-DNP antiserum.
`Experiments were in triplicate with average deviations given in parentheses.
`t Refers to concentration prior to the addition of goat anti-rabbit Fe antiserum.
`§ Expressed as percentagi; of the quantity precipitated in the absence of competitor.
`
`I- IV. Each table reports data obtained with a different donor-recipient pair of
`rabbits. The last column in each table presents relative binding affinities of
`haptens, obtained by Pressman et al. (9), \Yho measured inhibition-of specific
`precipitation by haptens.
`It is evident that specific haptens inhibit the reactions of anti-D sera. By far
`the best inhibitor in each system was p-(p'-hydroxy)-phenylazobenzoate. This
`
`PFIZER EX. 1103
`Page 15
`
`

`

`954
`
`QUANTITATIVE INVESTIGATIONS OF IDIOT YP IC ANTIBODIES. IV
`
`compou nd also combines with greater affi nity with anti-p-azobenzoate anti(cid:173)
`bodies than any of the other compounds tested. The percentage inhibi tion ob(cid:173)
`served in the four systems ranged from 43 to 69 % when p-(p'-hydroxy)-phenyl(cid:173)
`azobenzoate _was present at a final c.oncentration of l.6 X 10- 3 M.
`In tliree of the four systems . (Tables I- III), all para-substituted benzoate
`derivatives tested were quite effective as inhibitors, and were more effective
`th a