AD-R57 935 IMMUNOTECHNDLOGY(U) GEORGETON UNIV NASHINGTON D C i/i

UNCLASSIFIED F/G 6/5 N

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NATIONAL IWmEAU OF STANDADSMalOCOpy RESUCLTl TEST CHART

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* Ig~ECURITY CLASSIFICATION Ow 711611 PAGE (When Die sneers*iIIi~REPORT DOCUMENTATION PAGE 11FR OPEIGFR

11. REORT NUS94 GVT ACCESSION NO. S. 0SCIPIEgTi5 CATALOG NUMBER

in4M'W 4. TITLE re'd 8.ibthl) a. TYPE OF REPORT & PERIOD COVERED

(n mmuotehnoogyAnnual 10/31 83 to 10/31 84

N S.tcholg PERFORMING ONG. REORMUMOLK

t) 7.AUTHOn(e) 6.- 01TRACT 00 GRNT kumset(o)

Philip Lake, Ph.D. NOOO- 14-80-K-0909

9 . PERFORMING ORGANIZATION RNK AND ADRESS 10. PROGRAM ELEMENT PROJECT. TASK

Georgetown University Medical Center NR 666-003

*II. COWTNOLLIWG 00OPiCE AM a AONtSS 12. REPORT DATEJeannine A. Majde, Ph.D., Scientific Officer Oct. 31, 1984Immunology, Code 441, Cellular Biosystems Group IS. NUN9uER OF PAGESDept. of the Navy, ONR, Arlington, VA. '22217 16

14. MONITORING AGENCV NAME & ADDRESS(lI 011opent kes CentmiffM 0Offic) 16. SECUiITV CLASS. (*f this tpo'W)

Jeannine A. Majde, Ph.D., Scientific Officer UcasfeImmunology, Code 441, Cellular Biosystems Group UnclassifieAIIDONRDGDept. of the Navy, ONR, Arlington, VA. 22217 09a wausSPCtOf,6RN

SC ULE Unlimited14. OISTRIUIONa STATEMENT (ofOhi Rpe 31)

*17. OiSTRIOUTION ITATEMENS1T (of the abstract onteret in 85.65c 20. it Affeent 111pA Rop.,rt)

Unlimited

* IS. SUPPLEMENTARY NOTES

3 It. KEY WOODS 'Centle. on rere Olds 110080 dodem 11 dntitb by Week nmb.,)

La.J Gamma interferon, B cells, TRF T cell replacing factor, interleukin-1LAJ

C2 immunobiology. In this study we have examined the control of B cell responsesivitro by several lymphokines including IL-i, IL-2 and gamma interferon.

This analysis provides information on the potential to manipulate B cells inculture for their production of desired antibodies, as may be needed prior tohybridoma production and also provides information on the mechanism ofregulation of lymphocyte responses.

In the present study we have shown that IL-i and more importantly gammna

DO W 43 £,rt Or@I Nov so towo,LrTr

83 O)~LP @* O6 CURImT CLASSIFICATION Of THIS PAGE1 (manm beto "tepee)

a.. . . . . . .. . . . . . . .

SUCUPATV CLAShIFICATION OF THIS PAGE (111640e O@.Rf#

* interferon are involved in the regulation of B cell responses. The role ofgamma interferon in T cell-replacing factor (TRF) activity for antigen-specificplaque-forming cell (PFC) responses in vitro was studied using antibodies tomurine gamma interferon. TRF activity was present in supernatants (Sn) of ConA- or mixed leukocyte reaction-stimulated murine spleen cells as well as in anIL-2-rich fraction of phytohemagglutinin-stimulated murine human peripheral

-blood lymphocyte Sn and in the Sn of the Gibbon T lymphoma MLA-144. The human-TRF was highly active with cells from nu/nu mice and normal mice but not with

cells from animals with the xid immunologic defect, similar to the activity ofmurine TRF. 2(Antibodies to gamma interferon consisted of hyper-immune rabbitantisera, Gamma interferon affinity-purified rabbit immunoglobulin and aninterspectes hybridoma specific for murine gamma interferon. The results showthat the activities of all preparations of TRF are markedly diminished orabrogated by antibody to murine gamma interferon but not by antibodies to humangamma interferon, nor by normal rabbit sera or purified rabbit Ig. The degreeof inhibition was dose dependent and was quantitatively reversed by the

* addition to the cultures of recombinant-derived murine gamma interferon but not*human gamma interferon. This reversal was fully antigen specific and thus not

attributable to polyclonal B cell activation by gamma interferon, which isinactive alone in the TRF assay. Kinetic analysis shows that gamma interferonmust act by 24-48 h to produce PFC responses at 4 d. Together, the datademonstrate that gamma interferon is a necessary mediator for TRF effects andthat gamma interferon is induced by TRF from T-depleted murine spleen cells in

* sufficient quantity to support large antibody responses. The source of thisgamma interferon may be the potent natural killer cells that are induced incultures stimulated with TRF.

1

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FILMED

10-85

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