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Detection of genital tract sperm antibodies in systemically and locally immunized female rabbits
54
P 33
EXPERIMENTAL AL~OIMMUNE ORCHITIS IN WISTAR RATS. I~ STUDY OF LYMPHOCYTE SUBSETS AT THE IMMUNIZATION DRAINING LYMPH NODES. G.F. Doncel and L. LUSTIG, Centro de Investigaciones en Reproduccion, Facultad de Medicina, Universidad de Buenos Aires, 1121 Buenos Aires, Argentina. T and B cell populations at the immunization draining lymph nodes were stud~d by immunofluorescence using Mn Abs: W3/13(Pan T), W3/25(CD4), OX-8(CD8), OX-6 (MHC class II) and an anti-IgG,during the development of an autoimmune orchitis in Wistar rats. Animals immunized 3 times with a testicular homegenate (experimental group) or saline (control group) and Freund complete adjuvant and Bordetella Pertussis were sacrificed at 7,16,30,50 and 80 days,after the first immunization. Rats from control group showed a significant decrease of the CD4/ CD8 ratio, at 50 and 80 days(l.81±0.10 and 1.63± 0.i0) compared to non-immunized rats(2.72±0.18), basically due to an increase in CD8 + cells. In the experimental group, a testicular damage characterized by aspermatogenesis and orchitis appeared at 50 days in 60% of the rats. Normal percentages(CD4/CD8 ratio:2.63 ± 0.15), with an increase in the absolute cell n~mber of the different lymphocyte subsets were detected in animals with testicular pathology, while a significant augment of CD8 + cells(low ratio:l.92±0.18) was seen in rats without lesion.These rats also presented a higher level of W3/13 + cells and no significant differences in ~ - 6 + or B cells. Although the orchitis was more severe at 80 days, animals with or without lesion presented low ratios(l.95±0.12 and 1.77±0.07) due to an increase in CD8 + cells. In conclusion, the onset of testicular damage (50 days) correlated with a normal CD4/CD8 ratio in the lymph nodes, while the lack of lesion was concomitant with a "suppressive state" in them(high proportion of CD8+ cells. Keywords: autoimmunity; orchitisj lymphocyte subsets.
P 34 D E T E C T I O N OF G E N I T A L T R A C T S P E R ~ A N T I B O D I E S LOCALLY IMMUNIZED FEMALE RABBITS
IN S Y S T E M I C A L L Y
AND
M , D u b o v a , V , V u l c h a n o v ~ K , B r a t a n o v I n s t i t u t e of Biol. and Immunol. of P e ~ r o d . ~ Bulg. A c a d . Sci0~ 73 L e n i n B l v d . , l l l 3 S o f i a , B u l g a r i a ~ature female rabbits were immunized with washed sonified allogen e i c s p e r m a t o z o a i.d., i.v. o r / a n d i n t r a v a g i n a l l y (antigen being i n t r o d u c e d b y c a t h e t e r s or a p p l i e d i n c o r p o r a t e d in a g a r o s e gels). The i m m u n o g e n was p r o v e d to c o n t a i n m o r e t h a n 10 s o l u b l e a n t i g e n i c c o m p o n e n t s . The i m m u n e r e s p o n s e as e s t i m a t e d by w e e k l y E L I S A c h e c k s of s e r a , c e r v i c o v a g i n a l (c.v.) s e c r e t i o n s , f a l l o p i a n tube and u t e r i n e w a s h i n g s m a n i f e s t e d in s y s t e m i c i m m u n i z a t i o n h i g h t i t r e I~G a n d no I~A s p e r m a n t i b o d i e s (Ab). In g e n i t a l t r a c t s e c r e t i o n s of s y s t e m i c a l l y t r e a t e d a n i m a l s IgG s p e r m Ab w e r e e s t a b l i s h e d in rabbits showing s e r u m IgG t i t r e s a b o v e l : 4 0 0 . A t t e m p t s at i n d u c i n g local a n t i s D e r m response by v a g i n a l Ag a p p l i c a t i o n s h o w e d that o n l y m u l t i p l e (4-8 t i m e s x6 hrs) Ag a p p l i c a t i o n r e s u l t e d in the f o r m a t i o n o f s p e r m Ab, the r e s p o n s e b e i n g l o c a l and s y s t e m i c b u t n e v e r l o c a l alone. In c.v. s e c r e t i o n s of v a g i n a l l y t r e a t e d r a b b i t s IgA a n d I~G s p e r m Ab w e r e found. The a b s e n c e or the low t i t r e of I~A s n e r m Ab in the sera of the c o r r e s p o n d i n g f e m a l e s s u g g e s t s t h e i r l o c a l p r o d u c t i o n . O u r r e s u l t s s u g g e s t also that p a r t of IgG s p e r m Ab in c.v. s e c r e t i o n s of l o c a l l y t r e a t e d a n i m a l s c o u l d be of local o r i g i n too, b e c a u s e of the v e r y l o w s e r u m IgG l e v e l of s p e r m Ab in the c o r r e s p o n d i n g r a b b i t s . immunization,
local,
sperm
P 33
EXPERIMENTAL AL~OIMMUNE ORCHITIS IN WISTAR RATS. I~ STUDY OF LYMPHOCYTE SUBSETS AT THE IMMUNIZATION DRAINING LYMPH NODES. G.F. Doncel and L. LUSTIG, Centro de Investigaciones en Reproduccion, Facultad de Medicina, Universidad de Buenos Aires, 1121 Buenos Aires, Argentina. T and B cell populations at the immunization draining lymph nodes were stud~d by immunofluorescence using Mn Abs: W3/13(Pan T), W3/25(CD4), OX-8(CD8), OX-6 (MHC class II) and an anti-IgG,during the development of an autoimmune orchitis in Wistar rats. Animals immunized 3 times with a testicular homegenate (experimental group) or saline (control group) and Freund complete adjuvant and Bordetella Pertussis were sacrificed at 7,16,30,50 and 80 days,after the first immunization. Rats from control group showed a significant decrease of the CD4/ CD8 ratio, at 50 and 80 days(l.81±0.10 and 1.63± 0.i0) compared to non-immunized rats(2.72±0.18), basically due to an increase in CD8 + cells. In the experimental group, a testicular damage characterized by aspermatogenesis and orchitis appeared at 50 days in 60% of the rats. Normal percentages(CD4/CD8 ratio:2.63 ± 0.15), with an increase in the absolute cell n~mber of the different lymphocyte subsets were detected in animals with testicular pathology, while a significant augment of CD8 + cells(low ratio:l.92±0.18) was seen in rats without lesion.These rats also presented a higher level of W3/13 + cells and no significant differences in ~ - 6 + or B cells. Although the orchitis was more severe at 80 days, animals with or without lesion presented low ratios(l.95±0.12 and 1.77±0.07) due to an increase in CD8 + cells. In conclusion, the onset of testicular damage (50 days) correlated with a normal CD4/CD8 ratio in the lymph nodes, while the lack of lesion was concomitant with a "suppressive state" in them(high proportion of CD8+ cells. Keywords: autoimmunity; orchitisj lymphocyte subsets.
P 34 D E T E C T I O N OF G E N I T A L T R A C T S P E R ~ A N T I B O D I E S LOCALLY IMMUNIZED FEMALE RABBITS
IN S Y S T E M I C A L L Y
AND
M , D u b o v a , V , V u l c h a n o v ~ K , B r a t a n o v I n s t i t u t e of Biol. and Immunol. of P e ~ r o d . ~ Bulg. A c a d . Sci0~ 73 L e n i n B l v d . , l l l 3 S o f i a , B u l g a r i a ~ature female rabbits were immunized with washed sonified allogen e i c s p e r m a t o z o a i.d., i.v. o r / a n d i n t r a v a g i n a l l y (antigen being i n t r o d u c e d b y c a t h e t e r s or a p p l i e d i n c o r p o r a t e d in a g a r o s e gels). The i m m u n o g e n was p r o v e d to c o n t a i n m o r e t h a n 10 s o l u b l e a n t i g e n i c c o m p o n e n t s . The i m m u n e r e s p o n s e as e s t i m a t e d by w e e k l y E L I S A c h e c k s of s e r a , c e r v i c o v a g i n a l (c.v.) s e c r e t i o n s , f a l l o p i a n tube and u t e r i n e w a s h i n g s m a n i f e s t e d in s y s t e m i c i m m u n i z a t i o n h i g h t i t r e I~G a n d no I~A s p e r m a n t i b o d i e s (Ab). In g e n i t a l t r a c t s e c r e t i o n s of s y s t e m i c a l l y t r e a t e d a n i m a l s IgG s p e r m Ab w e r e e s t a b l i s h e d in rabbits showing s e r u m IgG t i t r e s a b o v e l : 4 0 0 . A t t e m p t s at i n d u c i n g local a n t i s D e r m response by v a g i n a l Ag a p p l i c a t i o n s h o w e d that o n l y m u l t i p l e (4-8 t i m e s x6 hrs) Ag a p p l i c a t i o n r e s u l t e d in the f o r m a t i o n o f s p e r m Ab, the r e s p o n s e b e i n g l o c a l and s y s t e m i c b u t n e v e r l o c a l alone. In c.v. s e c r e t i o n s of v a g i n a l l y t r e a t e d r a b b i t s IgA a n d I~G s p e r m Ab w e r e found. The a b s e n c e or the low t i t r e of I~A s n e r m Ab in the sera of the c o r r e s p o n d i n g f e m a l e s s u g g e s t s t h e i r l o c a l p r o d u c t i o n . O u r r e s u l t s s u g g e s t also that p a r t of IgG s p e r m Ab in c.v. s e c r e t i o n s of l o c a l l y t r e a t e d a n i m a l s c o u l d be of local o r i g i n too, b e c a u s e of the v e r y l o w s e r u m IgG l e v e l of s p e r m Ab in the c o r r e s p o n d i n g r a b b i t s . immunization,
local,
sperm