Adjuvants and aspermatogenesis

80 immuno-affinity column was prepared with IgG isolated from 8CI0.5 ascites and coupled to glutaraldehyde-activated agarose gel. A preparation of DOCsolubilized testis was run through the MAb affinity column and fractions eluted using pH in increasing steps (pH8-11.4). A purified antigen was isolated showing a single band of 63 kd in SDS-PAGE. The antigen which was identified as a glyco-protein was immunogenic in mice but not in rabbits. The murine antisperm (a/s) was monospeeific recognizing a single protein molecule in single dimensional and 2D gel/protein blot procedure reacting with specific peptides of differing charge but same molecular weight. The a/s which was sperm/testis specific did not agglutinate nor immobilize sperm but did react in immunofluorescence not only with rabbit sperm but also with human and murine sperm. Artificial insemination of female rabbits with a/s-treated sperm resulted in a significant inhibition of fertility at 9 days p.c. without affecting fertilization rates. The post fertilization antifertility effect of a/s was not due to parthenogenie activation of ova or to polyspermy. Passive transfer (i.p.) of antibodies into mice suggested that the anti-63kd antiserum reduced fetal survival. The immuno-contraceptive potential and immunoinfertility diagnostic values of the antigen are under study. PgE 2 M O D U L A T I N G

EFFECTS

IMMUNE

TO SPERM ANTIGENS

RESPONSE

ON THE CELL-MEDIATED

AND

HUMORAL

A.P.TORNYOV, V.H.VULCHANOV, B.A.VELEV I n s t i t u t e of B i o l o g y a n d I m m u n o l o g y of R e p r o d u c t i o n , A c a d e m y of S c i e n c e s , S o f i a , B u l g a r i a

Bulgarian

The i.[). i n t r o d u c t i o n of P r o s t a g l a n d i n Ep (PgEg) in B a l b / c m i c e delays" the r e j e c t i o n of C 5 7 B L 6 - m i c e - g r a f [ s w h i l e the i n j e c t i o n of I n d o m e t h a c i n u m / t h e p r o s t a g l a n d i n - s y n t h e t h a s e inhibitor/ a c c e l e r a t e s t h i s p r o c e s s . The s a m e t e n d e n c i e s are m a r k e d a l s o in the 2 n d set r e a c t i o n , as w e l l in the c a s e s w i t h t r a n s p l a n t a t i o n of m a l e B a l b / c s k i n on f e m a l e s of t h e same s t r a i n / H - Y g e n e t i c d i f f e r e n c e s / . It is n o t a b l e a l s o t h a t B a l b / c - m i c e w h i c h r e c e i v e PgE~ during their immunization with epidydimal spermatozoa from C 5 7 B L 6 r e j e c t m o r e s l o w l y s k i n g r a f t s f r o m the d o n o r s t r a i n in c o m p a r i s o n w i t h m i c e i n j e c t e d w i t h s p e r m a t o z o a only. D a t a s h o w i n g immunosuppressive a c t i o n of P g E 2 on the p r i m a r y a n d s e c o n d a r y h u m o r a l i m m u n e r e s p o n s e to S R B C or to a l l o g e n i c a n d h e t e r o g e n i c spermatozoa were also obtained. A h y p o t h e s i s for the i m m u n o r e g u l a t o r y r o l e of P g E 2 in the d e t e r m i n a t i o n of n a t u r a l i m m u n o l o g i c a l t o l e r a n c e to s p e r m a n t i g e n s in the g e n i t a l t r a c t is b e i n g p r o p o s e d .

ADJUVA}]TS AND ASPEP~MATOGENE,CIS

SEYMOUR KATSH AND GRACE F. KATSI!. DEPARTMENT OF PHA.~MACOLOGY U~IVERSITY OF COLORADO SCIIOOL OF MEDICINE, DE~ER, COLORADO 80262, U..C.A. A single injection of specific antigen (from homologous or autologous sperm, testis or epididymis) when incorporated in appropriate a~juvant, induces deletion of the seminiferous epithelium in the guinea pi~. ['~ile the Freund adjuvant is effective, it is not acceptable for use in humans. After several hundred trials with oils, emulsifiers and bacterial fractions (c.f. Int. Arch. Allergy & Im~unol., 15:172, 1959; idem, 24:319, 1064; i~ature 212:1486, i ~ 6 6 ; Life Sciences, 201:761, 1977) we now find that the mineral oil component can be

8! replaced with certain plant oils, (jojoba oil is one of the most effective) and, the mycobacterial component can be replaced with purified fractions from Corynebacterium rubrum as well as the synthetic peptide N-acetyl-muramyl--L-alaD-GIu-Nii~ in ug. amounts. ~mlongst the ineffective adjuvants were Evans Blue (T-1824)~and sodium phthalyl lipo-polysaccharide (E. Coli K235). Adjuvants effective in inducing high humoral antibody are not able to induce aspermatogenesis in a single injection. These studies are important not only in immuno-reproduction but also for immuno-oncology. As a result of these series of studies, we believe an adjuvant compatible with human use has been constructed. (Supported by grants from the NSF, the Population Council, the NIH and the Ford Foundation). ~0i',i~ A ~ I ~ C T S

OF Ii~.,~UhOGLNIC

~STLIIILITY

OP AN I M A L S EUGENE

ILYI$LKY

USSR,

350044,

~'he p o s s i b i l i t y observed monas

in bulls

occered.

moantibodies the

sterility

for

the

even

the

spontaneous

using

of

the

the

KALININ

stimulus

INSTITUTE.

ST.,

I3.

autospermoantibodies and

experimental

bulls'

fertilisation

if

AGI{ICULTURAL

KRASNODAR,

of m a k i n g

when

The

,KUBAN

of

sperm the

with

cows

pseudomonas

was

pseudoautosper-

carried had

to

influence

on n e g a t i v e l y . The titis ral

is

breaking explained

regulation

mation

of

structure

of

the of

of

the

not the

cows'

only

the

sexual

and

the

changes

processes,

spermoantibodies sperme

reproduction

during

of

the n e u r o - h u m o -

but

associated

pathologically

the m a s -

also

with

by

the

for-

antigenic

changes

of m i l k .

INDUCTION, IMMUNOPATHOLOGY AND PATHOGENESIS OF MURINE EXPERIMENTAL ALLERGIC ORCHITIS (EAO). K.S.K. Tung, C. Teuscher, Suzanne M. Smith, T. Williams, J. Huser and S. Kohno. Dept. Pathology, Univ. New Mexico, Albuquerque, NM, USA EAO was induced consistently in Balb/c (H-2 d) mice by immunization with homologous testis homogenate in CFA and i ~g of B. pertussis extract (Infec. Immun.3:495, 1981). All animals developed orchitis and/or vasitis 20 day~ later. Murine EAObiS controlled, in part, by MHC genes since Balb. K (H-2~) and Balb. B10 (H-2) mice did not develop orchitis. Furthermore, orchitis and vasitis are probably dissociable since some mouse strains (SJL, NZB, C57BI/6) presented mainly orchitis, while other strains (Balb/c. Ig-c, Balb/c. Ig-d and Balb. K) developed mainly vasitis. By immunofluorescence, Ig was found in, or adjacent to Sertoli cells which showed degenerative changes on electron microscopy. The transitory changes, which occurred before EAO onset, are anti-