Vol. March
183,
No.
2, 1992
BIOCHEMICAL
AND
BIOPHYSICAL
RESEARCH
COMMUNICATIONS
Pages 538-543
16, 1992
WITH
INFERTILITY SYNTHETIC
INDUCED IN RATS BY IMMUNIZATION PEPTIDE SEGMENTS OF A SPERM PROTEIN
G. Vanage’, Y.A. Lu2, J.P. Tam2 and S.S. Koidel* 1 Center for Biomedical Research, The Population Council, New York, New York 1002 1 2The Rockefeller Received
January
24,
University,
New York, New York 1002 1
1992
SUMMARY: Three peptide segments(YAL-198, YAL-201 and YAL-212) corresponding to the extracellular domain of a humanspermprotein designatedas YWK-II antigen were synthesized asmultiple antigenpeptide (MAP). Male and female rats were immunized with the YWK-II-MAPS and fertility determined. In a group of 12 female rats immunized with YAL-198, seven animalswere infertile and two animalswere subfertile. When immunized with YAL-201 and YAL-212, 4 and 2 animalswere infertile, respectively. In a group of 15 malesimmunized with YAL-1982 animalswere infertile and 6 were subfertile. Two animals immunized with YAL-201 were subfertile. All control male and female rats immunized with bovine serumalbumin and adjuvant were fertile. Sera obtained from infertile rats immunized with YAL-198 contained higher titers of antibodiescomparedto thoseobtained from fertile animals. The presentstudy showsthat immunization with synthetic peptide segmentsof a spermprotein can effectively reduce fertility. a 1992?.ca&?mrc pre**, 1°C.
Infertility of undeterminedetiology is a clinical entity of considerablemagnitude that requires resolution (1). “Immunologic infertility” is due to the production of antigamete antibodies, derangementof B and/or T cell activities in the reproductive tract or alteration in the functional components of the immune network (2-6). There is a consensusthat antispermantibodies with spermagglutinating and/or immobilizing activities may be a probable cause of infertility; however, direct experimental verification of this thesis is lacking. There are studiessupporting this contention. For example, immunization of animals with sperm, sperm extracts or purified spermproteins will result in a marked reduction of fertility, suggestingthat production of antispenn antibodies can reduce fertility or induce infertility (7- 14). To validate that antispermantibodieswith spermagglutinating activity is a causeof infertility, we have raised monoclonal antibodies (mAb) against specific human sperm proteins with spermagglutinating activity (15-17). One of the antispermmAb designated *To whom correspondenceshouldbe addressed. 0006-291X192 $1.50 Copyright 0 1992 by Academic Press. Inc. All rights of reproduction in any form reserved.
538
Vol.
183, No. 2, 1992
BIOCHEMICAL
AND BIOPHYSICAL
RESEARCH COMMUNICATIONS
as YWK-II induced agglutination of humanand rat sperm(18-20) and prevented fertilization of zona-free hamstereggsby human sperm(21). Using an immunocytochemical method, the interacting antigen was located at the equatorial region of the head, midpiece and tail of human sperm(19). The equatorial region of the spermis purported to be the site of sperm-egg interaction during fertilization (22,23). In the rat, this protein is present on spermhead and produced in all germ cells undergoingspermatogenesis (20). A 1.8 kb cDNA for the YWK-II protein was isolated from a rat testis ilgtl 1 cDNA expressionlibrary and its nucleotide sequencedetermined(24). The cDNA contained an open reading frame coding for 191 amino acid residues. The deducedpolypeptide had high homology to the transmembrane-cytoplasmicdomainsof the A4 amyloid precursor protein found in brain plaquesof Alzheimer’s disease. The extracellular domain of the deduced YWK-II polypeptide, however, was unique. In the presentstudy, peptide segments correspondingto extracellular domain of the YWK-II protein were synthesized as multiple antigen peptide (MAP) (25,26). Data are presentedshowing that immunization of adult female rats with YWK-II-MAPS induced infertility in a significant number of animals. EXPERIMENTAL
METHODS
Synthesisof YWK-II-MAP Three peptide segmentscorrespondingto the extracellular domain of the YWK-II polypeptide were synthesizedasMAPS: YAL- 198, residue2 l-36, SEEIPPFHPFHPFPSL-Y; YAL-201, residue24-47, IPPFHPFHPFPSLSENEDTQPELY; and YAL-212, residue 5-36, SSISENPVDVRVSSEESEEIPPFHPFHPFPSL-Y. The synthesiswas performed asdescribedby Tam (25) The peptidesobtained from the synthesiswere dialyzed using a Spectra Por 6 tubing with Mr cut off of 1000 against a solution of 2 M urea, 0.1 M NH,HCO,! 0.01 M NH+,HCO, and water. The MAPS were lyophilized in water. Amino acid analysiswasperformed in 5.7 N HCl for 24 h at 110°C. Amino acid analysisshowedthat the compositioncorrespondsto the theoretical ratio.
Immunization procedure andfertility assay Groups of 15 male and 12 female adult rats were immunized with 100 pg of bovine serum albumin (control) and YWK-II-MAPS with MDP-P-T as adjuvant (27) at 2 week intervals over a 12 week period. Immunized female rats were mated with proven fertile male rats one week after the last immunization. Immunized male rats were mated with fertile female rats. The next day after copulation, the vagina was examined for sperm. On day 14 of expected pregnancy the animals were killed and uterus examined foi embryos. The number of embryos in each horn was counted. All control animalswere fertile containing 12 to 15 embryos in the uterus. Animals were consideredas infertile when no embryo was found and subfertile or reduced fertility when there were 6 to 9 embryos per uterus.
Assay of antibody titers MAPS were radioiodinated with “‘1 usingmodified chloramine T method b? MidgIcy (28). The circulating antibody titre was expressedaspercent binding of [I2 IlMAPs with individual serumdiluted to 1:50, determinedby radioimmunoassay. 539
Vol.
183,
No.
2, 1992
BIOCHEMICAL
AND
BIOPHYSICAL
RESEARCH
COMMUNICATIONS
RESULTS Groups of 12 female rats were immunized with YAL-198, YAL-201 and YAL 242 and their fertility determined (Table I). On immunization with YAL-198,7
animals were
infertile and 2 were subfertile. The uteri of the infertile animals were devoid of embryos (Fig. 1). Immunization rats, respectively.
with YAL-201 and YAL-212 induced infertility in 4 and 2 female
In male rats, immunization
with YAL-198 resulted in 2 infertile and 5
subfertile animals out of a total of 15 animals (Table II). Two male rats immunized with YAL-201
were subfertile
YWK-II-MAPS
and with YAL-212
none were affected (Table II).
tested, YAL-198 was the most effective in inducing infertility
Of the
in female
and male rats. The antibody titers in sera obtained from YWK-II-MAPS-treated
male and female
rats were determined by radioimmunoassay. The titers are expressed as percent binding at 1:25 and 1:50 dilution of male and female sera, respectively. Sera obtained from the seven infertile, two subfertile and three fertile females immunized with YAL-198 (Table I) showed binding titers ranging from 41 to 52%, 11 and 22%, and 3 to 18%, respectively. Infertile female rats immunized with YAL-201 and YAL-212 showed antibody binding titers of 30-40% and 25-30%, respectively, and fertile females, 5-20%. Sera (1:25 dilution) obtained from the two infertile males immunized with YAL-198 (Table II) showed antibody titers of 40 and 48% binding; five subfertile males, 15 to 30%; and eight fertile males, 4 to 20%. The binding titers of sera obtained from the two subfertile male rats immunized with YAL-201 (Table II) were 22 and 25%. Those obtained from the fertile male rats immunized with YAL-201 and YAL-212 were 5-20%. The present results suggest that infertility
may be related to circulating antibody titers.
DISCUSSION The present study is the first demonstration that immunization with synthetic peptide segments of a sperm protein can cause infertility.
This finding suggests that peptide
Table I. Fertility of adult female rats immunized with YWK-II-MAP No. of animals Status of fertility
YAL- 198
YAL-201
Infertile*
7
Subfertile+ Normal Pregnancy
2 3
4 0 8
YAL-212 2 0 10
Control female rats were immunized with bovine serum albumin and adjuvant; all rats were pregnant with 15 to 20 embryos/animal. *Infertile, no embryos, non-pregnant. +Subfertile, reduced no. of embryos: 6 to 9 embryos/animal. 540
Vol.
183, No. 2, 1992
BIOCHEMICAL
AND BIOPHYSICAL
RESEARCH COMMUNICATIONS
u. Uteri of female rats immunized with YAL- 198. Specimens were dissected out on day 14 after successful copulation. Lower, from infertile rats; upper. from a control fertile rat.
segmentscorresponding to epitopes or domain of specific spermprotein can be used as antifertility immunogens. The whole intact protein is not required. It should be pointed out that although YAL-198 was the most efficacious immunogentested, the longer segment, YAL-212 containing the samepeptide sequenceas YAL 198 was lesseffective. This finding that the antifertility potency waslesswith the longer segmentis not clear. A possiblereason is that YAL-198 contains the critical region for sperm-egginteraction,
Table II. Fertility of adult male rats immunized with YWK-II-MAP No. of animals Status of fertility
YAL- 198
YAL-20 1
2 5 8
0 2
Infertile* Subfertile+ Fertile Control male rats were immunized
13
with bovine serum albumin and adjuvant.
fertile and produced normal pregnancy in the females (15 to 20 embryos/animal). *Infertile, no embryos, non-pregnant. +Subfertile, reduced number of embryos, 6 to 9 embryos/animal.
541
YAL-212 0 0 15 All were
Vol.
183,
No.
2, 1992
BIOCHEMICAL
AND
whereas in the longer peptide, YAL-212,
BIOPHYSICAL
RESEARCH
COMMUNICATIONS
the extra peptide segment may reduce the
antigenicity by structural hindrance that the raised antibodies are less effective preventing sperm-egg interaction.
in
Polyclonal antibodies were raised against YAL- 198 in rabbits. The antisera induced tail-to-tail
agglutination of rat and human sperm and stained a 60-kD human sperm pro-
tein by immunoblot YWK-II
(unpublished data). In a previous study, it was demonstrated that
mAb interacted with a 60-kD human sperm protein and induced agglutination of
human sperm (17). Thus the above finding indicates that the YAL-198 correspond to a segment present in the 60-kD sperm protein There are several reports showing that fertility sperm proteins. infertility
Immunization
is reduced by immunization
with
with a sperm protein designated as PH-20 induced 100%
in guinea pigs (14). A sperm specific glycoprotein
designated as FA-1 from
human and mouse germ cell plasma membranes causes reduction of fertility in actively immunized female rabbits (13). Immunization of female rats with a 24 kD protein isolated from rat testicular cytosol induced infertility
in over 80% of treated animals (29).
Another soluble antigen, LDH-C,, is effective as antifertility immunogen (30). In these reported studies purified proteins were used; whereas in the present study synthetic peptide segments of known
sequence were utilized as immunogens and were effective in
reducing fertility. In summary, active immunization sperm protein can induce infertility YWK-II
with synthetic peptide segments of a specific
and/or reduced fertility in male and female rats. The
MAP designated as YAL- 198 should be further evaluated as a potential antifer-
tility vaccine. Acknowledgments: Rockefeller
This study was supported by grants from the Mellon Foundation and
Foundation and NIH AI 28701.
REFERENCES 1. Aral, S.O. and Cates, Jr., W. (1983) JAMA 250,2327-233 1.
2. Jones, W.R. (1980) Fertil. SteriZ. 33,577-586. 3. Haas, Jr., G.G., Cines, D.B., and Schreiber, A.D. 303,722-727.
(1980) N. Eng. J. Med.
4. Bronson, R.G., Cooper, G., and Rosenfeld, D. (1984) FertiZ. Steril. 42,171-182. 5. Shulman, S. (1986) Am. J. Reprod. Immunol. Microbial. 10,86-89. 6. Hargreave, T.B. (1982) Int. J. Fertil. 27,90-94. 7. Katsh, S. (1959) Am. J. Obstet. Gynecol. 78,276-278. 8. MC Laren, A. (1964) Nature 201,582-585. 9. Edwards, R.G. (1964) Nature 203,50-53. 542
Vol.
183,
No.
BIOCHEMICAL
2, 1992
AND
BIOPHYSICAL
RESEARCH
COMMUNICATIONS
10. Tyler, A. (1961) J. Reprod. Fertil. 2,473-506. 11. Syner, F.N., Kuros, B.S., and Moghissi, K.S. (1979) Fertil.
Steril. 32,468-473.
12. Tung, K.S.K., Goldberg, E.H., and Goldberg, E. (1979) J. Reprod.
Immunol.
1,145158. 13.
Naz, R.K. (1987) J. Reprod. Zmmunol. 11 ,I 17-133.
14. Primakoff, P., Lathop, W., Woolman, L., Cowan, A., and Myles, D. (1988) Nature 335543-546. 15. Yan, Y.C., Wang, L.F., Sato, E., and Koide, S.S. (1983) Am. .I. Reprod. lmmunol. 4,111-115. 16. Yan, Y.C., Mitsudo, S.M., Wang, L.F., and Koide, S.S. (1984) 42,614-617.
Fertil.
Steril.
17. Yan, Y.C., Wang, L.F., and Koide, S.S. (1986) Int. J. Fertil. 31,77-85. 18. Yan, Y.C., Wang, L.F., Mitsudo, S.M., and Koide, S.S. (1986) in fmmunological Approach to Contraception and Promotion of Fertility, G.P. Talwar, Eds, Plenum Publishing Corporation, 23l-240. 19. Yan, Y.C., Wang, L.F., and Koide, S.S. (1987) Arch. Androl.
18,245-254.
20.
HaneJi,T. and Koide, S.S. (1987) Biol. Reprod. 37,467-477.
21.
Wang, L.F., Yan, Y.C., Miao, S.Y., and Koide, S.S. (1987) in New Horizons Spermatozoa
Research,
in
H. Mohri. Eds, Japan Scientific Societies Press,Tokyo,
409-420. 22.
Shalgi, R. and Phillips, D. (1980) 1. Ultrustruct.
Res. 7 1,154- 161.
23.
Bedford, J.M., Moore, H.D.M., and Franklin, L.E. (1980) E.xp. Cell
Res.
119.119-126. 24.
Yan, Y.C., Bai, Y., Wang, L.F., Miao, S.-Y., and Koide, S.S. (1990) Proc. Natl. Acad. Sci. USA 87,2405-2408.
25.
Tam, J.P. (1988) Proc. Natl. Acad. Sci. USA 855409-5413.
26.
Tam, J.P. and Lu, Y.A. (1989) Proc. Natl. Acad. Sci. USA 86,9084-9088.
27.
Ladd, A., Gopinath, P., Tsong, Y.-Y., Probst, T., Chung, W., and Thau, R.B. (1988) Am. J. Reprod. Immunol. Microbial. 17,121-127.
28.
Midgley, Jr., A.R. (1966) Endocrinology
29.
Shaha, C., Suri, A., and Talwar, G.P. (1990) lnternat
30.
Goldberg, E. and Shelton, J.A. (1985) in lmmurwlogical ception
and Fertility,
79,10-18. J. Androl.
13,17-25.
Approaches
to Contra-
G.P. Talwar, Eds, PlenumPress,New York, 219-230.
543
183,
No.
2, 1992
BIOCHEMICAL
AND
BIOPHYSICAL
RESEARCH
COMMUNICATIONS
Pages 538-543
16, 1992
WITH
INFERTILITY SYNTHETIC
INDUCED IN RATS BY IMMUNIZATION PEPTIDE SEGMENTS OF A SPERM PROTEIN
G. Vanage’, Y.A. Lu2, J.P. Tam2 and S.S. Koidel* 1 Center for Biomedical Research, The Population Council, New York, New York 1002 1 2The Rockefeller Received
January
24,
University,
New York, New York 1002 1
1992
SUMMARY: Three peptide segments(YAL-198, YAL-201 and YAL-212) corresponding to the extracellular domain of a humanspermprotein designatedas YWK-II antigen were synthesized asmultiple antigenpeptide (MAP). Male and female rats were immunized with the YWK-II-MAPS and fertility determined. In a group of 12 female rats immunized with YAL-198, seven animalswere infertile and two animalswere subfertile. When immunized with YAL-201 and YAL-212, 4 and 2 animalswere infertile, respectively. In a group of 15 malesimmunized with YAL-1982 animalswere infertile and 6 were subfertile. Two animals immunized with YAL-201 were subfertile. All control male and female rats immunized with bovine serumalbumin and adjuvant were fertile. Sera obtained from infertile rats immunized with YAL-198 contained higher titers of antibodiescomparedto thoseobtained from fertile animals. The presentstudy showsthat immunization with synthetic peptide segmentsof a spermprotein can effectively reduce fertility. a 1992?.ca&?mrc pre**, 1°C.
Infertility of undeterminedetiology is a clinical entity of considerablemagnitude that requires resolution (1). “Immunologic infertility” is due to the production of antigamete antibodies, derangementof B and/or T cell activities in the reproductive tract or alteration in the functional components of the immune network (2-6). There is a consensusthat antispermantibodies with spermagglutinating and/or immobilizing activities may be a probable cause of infertility; however, direct experimental verification of this thesis is lacking. There are studiessupporting this contention. For example, immunization of animals with sperm, sperm extracts or purified spermproteins will result in a marked reduction of fertility, suggestingthat production of antispenn antibodies can reduce fertility or induce infertility (7- 14). To validate that antispermantibodieswith spermagglutinating activity is a causeof infertility, we have raised monoclonal antibodies (mAb) against specific human sperm proteins with spermagglutinating activity (15-17). One of the antispermmAb designated *To whom correspondenceshouldbe addressed. 0006-291X192 $1.50 Copyright 0 1992 by Academic Press. Inc. All rights of reproduction in any form reserved.
538
Vol.
183, No. 2, 1992
BIOCHEMICAL
AND BIOPHYSICAL
RESEARCH COMMUNICATIONS
as YWK-II induced agglutination of humanand rat sperm(18-20) and prevented fertilization of zona-free hamstereggsby human sperm(21). Using an immunocytochemical method, the interacting antigen was located at the equatorial region of the head, midpiece and tail of human sperm(19). The equatorial region of the spermis purported to be the site of sperm-egg interaction during fertilization (22,23). In the rat, this protein is present on spermhead and produced in all germ cells undergoingspermatogenesis (20). A 1.8 kb cDNA for the YWK-II protein was isolated from a rat testis ilgtl 1 cDNA expressionlibrary and its nucleotide sequencedetermined(24). The cDNA contained an open reading frame coding for 191 amino acid residues. The deducedpolypeptide had high homology to the transmembrane-cytoplasmicdomainsof the A4 amyloid precursor protein found in brain plaquesof Alzheimer’s disease. The extracellular domain of the deduced YWK-II polypeptide, however, was unique. In the presentstudy, peptide segments correspondingto extracellular domain of the YWK-II protein were synthesized as multiple antigen peptide (MAP) (25,26). Data are presentedshowing that immunization of adult female rats with YWK-II-MAPS induced infertility in a significant number of animals. EXPERIMENTAL
METHODS
Synthesisof YWK-II-MAP Three peptide segmentscorrespondingto the extracellular domain of the YWK-II polypeptide were synthesizedasMAPS: YAL- 198, residue2 l-36, SEEIPPFHPFHPFPSL-Y; YAL-201, residue24-47, IPPFHPFHPFPSLSENEDTQPELY; and YAL-212, residue 5-36, SSISENPVDVRVSSEESEEIPPFHPFHPFPSL-Y. The synthesiswas performed asdescribedby Tam (25) The peptidesobtained from the synthesiswere dialyzed using a Spectra Por 6 tubing with Mr cut off of 1000 against a solution of 2 M urea, 0.1 M NH,HCO,! 0.01 M NH+,HCO, and water. The MAPS were lyophilized in water. Amino acid analysiswasperformed in 5.7 N HCl for 24 h at 110°C. Amino acid analysisshowedthat the compositioncorrespondsto the theoretical ratio.
Immunization procedure andfertility assay Groups of 15 male and 12 female adult rats were immunized with 100 pg of bovine serum albumin (control) and YWK-II-MAPS with MDP-P-T as adjuvant (27) at 2 week intervals over a 12 week period. Immunized female rats were mated with proven fertile male rats one week after the last immunization. Immunized male rats were mated with fertile female rats. The next day after copulation, the vagina was examined for sperm. On day 14 of expected pregnancy the animals were killed and uterus examined foi embryos. The number of embryos in each horn was counted. All control animalswere fertile containing 12 to 15 embryos in the uterus. Animals were consideredas infertile when no embryo was found and subfertile or reduced fertility when there were 6 to 9 embryos per uterus.
Assay of antibody titers MAPS were radioiodinated with “‘1 usingmodified chloramine T method b? MidgIcy (28). The circulating antibody titre was expressedaspercent binding of [I2 IlMAPs with individual serumdiluted to 1:50, determinedby radioimmunoassay. 539
Vol.
183,
No.
2, 1992
BIOCHEMICAL
AND
BIOPHYSICAL
RESEARCH
COMMUNICATIONS
RESULTS Groups of 12 female rats were immunized with YAL-198, YAL-201 and YAL 242 and their fertility determined (Table I). On immunization with YAL-198,7
animals were
infertile and 2 were subfertile. The uteri of the infertile animals were devoid of embryos (Fig. 1). Immunization rats, respectively.
with YAL-201 and YAL-212 induced infertility in 4 and 2 female
In male rats, immunization
with YAL-198 resulted in 2 infertile and 5
subfertile animals out of a total of 15 animals (Table II). Two male rats immunized with YAL-201
were subfertile
YWK-II-MAPS
and with YAL-212
none were affected (Table II).
tested, YAL-198 was the most effective in inducing infertility
Of the
in female
and male rats. The antibody titers in sera obtained from YWK-II-MAPS-treated
male and female
rats were determined by radioimmunoassay. The titers are expressed as percent binding at 1:25 and 1:50 dilution of male and female sera, respectively. Sera obtained from the seven infertile, two subfertile and three fertile females immunized with YAL-198 (Table I) showed binding titers ranging from 41 to 52%, 11 and 22%, and 3 to 18%, respectively. Infertile female rats immunized with YAL-201 and YAL-212 showed antibody binding titers of 30-40% and 25-30%, respectively, and fertile females, 5-20%. Sera (1:25 dilution) obtained from the two infertile males immunized with YAL-198 (Table II) showed antibody titers of 40 and 48% binding; five subfertile males, 15 to 30%; and eight fertile males, 4 to 20%. The binding titers of sera obtained from the two subfertile male rats immunized with YAL-201 (Table II) were 22 and 25%. Those obtained from the fertile male rats immunized with YAL-201 and YAL-212 were 5-20%. The present results suggest that infertility
may be related to circulating antibody titers.
DISCUSSION The present study is the first demonstration that immunization with synthetic peptide segments of a sperm protein can cause infertility.
This finding suggests that peptide
Table I. Fertility of adult female rats immunized with YWK-II-MAP No. of animals Status of fertility
YAL- 198
YAL-201
Infertile*
7
Subfertile+ Normal Pregnancy
2 3
4 0 8
YAL-212 2 0 10
Control female rats were immunized with bovine serum albumin and adjuvant; all rats were pregnant with 15 to 20 embryos/animal. *Infertile, no embryos, non-pregnant. +Subfertile, reduced no. of embryos: 6 to 9 embryos/animal. 540
Vol.
183, No. 2, 1992
BIOCHEMICAL
AND BIOPHYSICAL
RESEARCH COMMUNICATIONS
u. Uteri of female rats immunized with YAL- 198. Specimens were dissected out on day 14 after successful copulation. Lower, from infertile rats; upper. from a control fertile rat.
segmentscorresponding to epitopes or domain of specific spermprotein can be used as antifertility immunogens. The whole intact protein is not required. It should be pointed out that although YAL-198 was the most efficacious immunogentested, the longer segment, YAL-212 containing the samepeptide sequenceas YAL 198 was lesseffective. This finding that the antifertility potency waslesswith the longer segmentis not clear. A possiblereason is that YAL-198 contains the critical region for sperm-egginteraction,
Table II. Fertility of adult male rats immunized with YWK-II-MAP No. of animals Status of fertility
YAL- 198
YAL-20 1
2 5 8
0 2
Infertile* Subfertile+ Fertile Control male rats were immunized
13
with bovine serum albumin and adjuvant.
fertile and produced normal pregnancy in the females (15 to 20 embryos/animal). *Infertile, no embryos, non-pregnant. +Subfertile, reduced number of embryos, 6 to 9 embryos/animal.
541
YAL-212 0 0 15 All were
Vol.
183,
No.
2, 1992
BIOCHEMICAL
AND
whereas in the longer peptide, YAL-212,
BIOPHYSICAL
RESEARCH
COMMUNICATIONS
the extra peptide segment may reduce the
antigenicity by structural hindrance that the raised antibodies are less effective preventing sperm-egg interaction.
in
Polyclonal antibodies were raised against YAL- 198 in rabbits. The antisera induced tail-to-tail
agglutination of rat and human sperm and stained a 60-kD human sperm pro-
tein by immunoblot YWK-II
(unpublished data). In a previous study, it was demonstrated that
mAb interacted with a 60-kD human sperm protein and induced agglutination of
human sperm (17). Thus the above finding indicates that the YAL-198 correspond to a segment present in the 60-kD sperm protein There are several reports showing that fertility sperm proteins. infertility
Immunization
is reduced by immunization
with
with a sperm protein designated as PH-20 induced 100%
in guinea pigs (14). A sperm specific glycoprotein
designated as FA-1 from
human and mouse germ cell plasma membranes causes reduction of fertility in actively immunized female rabbits (13). Immunization of female rats with a 24 kD protein isolated from rat testicular cytosol induced infertility
in over 80% of treated animals (29).
Another soluble antigen, LDH-C,, is effective as antifertility immunogen (30). In these reported studies purified proteins were used; whereas in the present study synthetic peptide segments of known
sequence were utilized as immunogens and were effective in
reducing fertility. In summary, active immunization sperm protein can induce infertility YWK-II
with synthetic peptide segments of a specific
and/or reduced fertility in male and female rats. The
MAP designated as YAL- 198 should be further evaluated as a potential antifer-
tility vaccine. Acknowledgments: Rockefeller
This study was supported by grants from the Mellon Foundation and
Foundation and NIH AI 28701.
REFERENCES 1. Aral, S.O. and Cates, Jr., W. (1983) JAMA 250,2327-233 1.
2. Jones, W.R. (1980) Fertil. SteriZ. 33,577-586. 3. Haas, Jr., G.G., Cines, D.B., and Schreiber, A.D. 303,722-727.
(1980) N. Eng. J. Med.
4. Bronson, R.G., Cooper, G., and Rosenfeld, D. (1984) FertiZ. Steril. 42,171-182. 5. Shulman, S. (1986) Am. J. Reprod. Immunol. Microbial. 10,86-89. 6. Hargreave, T.B. (1982) Int. J. Fertil. 27,90-94. 7. Katsh, S. (1959) Am. J. Obstet. Gynecol. 78,276-278. 8. MC Laren, A. (1964) Nature 201,582-585. 9. Edwards, R.G. (1964) Nature 203,50-53. 542
Vol.
183,
No.
BIOCHEMICAL
2, 1992
AND
BIOPHYSICAL
RESEARCH
COMMUNICATIONS
10. Tyler, A. (1961) J. Reprod. Fertil. 2,473-506. 11. Syner, F.N., Kuros, B.S., and Moghissi, K.S. (1979) Fertil.
Steril. 32,468-473.
12. Tung, K.S.K., Goldberg, E.H., and Goldberg, E. (1979) J. Reprod.
Immunol.
1,145158. 13.
Naz, R.K. (1987) J. Reprod. Zmmunol. 11 ,I 17-133.
14. Primakoff, P., Lathop, W., Woolman, L., Cowan, A., and Myles, D. (1988) Nature 335543-546. 15. Yan, Y.C., Wang, L.F., Sato, E., and Koide, S.S. (1983) Am. .I. Reprod. lmmunol. 4,111-115. 16. Yan, Y.C., Mitsudo, S.M., Wang, L.F., and Koide, S.S. (1984) 42,614-617.
Fertil.
Steril.
17. Yan, Y.C., Wang, L.F., and Koide, S.S. (1986) Int. J. Fertil. 31,77-85. 18. Yan, Y.C., Wang, L.F., Mitsudo, S.M., and Koide, S.S. (1986) in fmmunological Approach to Contraception and Promotion of Fertility, G.P. Talwar, Eds, Plenum Publishing Corporation, 23l-240. 19. Yan, Y.C., Wang, L.F., and Koide, S.S. (1987) Arch. Androl.
18,245-254.
20.
HaneJi,T. and Koide, S.S. (1987) Biol. Reprod. 37,467-477.
21.
Wang, L.F., Yan, Y.C., Miao, S.Y., and Koide, S.S. (1987) in New Horizons Spermatozoa
Research,
in
H. Mohri. Eds, Japan Scientific Societies Press,Tokyo,
409-420. 22.
Shalgi, R. and Phillips, D. (1980) 1. Ultrustruct.
Res. 7 1,154- 161.
23.
Bedford, J.M., Moore, H.D.M., and Franklin, L.E. (1980) E.xp. Cell
Res.
119.119-126. 24.
Yan, Y.C., Bai, Y., Wang, L.F., Miao, S.-Y., and Koide, S.S. (1990) Proc. Natl. Acad. Sci. USA 87,2405-2408.
25.
Tam, J.P. (1988) Proc. Natl. Acad. Sci. USA 855409-5413.
26.
Tam, J.P. and Lu, Y.A. (1989) Proc. Natl. Acad. Sci. USA 86,9084-9088.
27.
Ladd, A., Gopinath, P., Tsong, Y.-Y., Probst, T., Chung, W., and Thau, R.B. (1988) Am. J. Reprod. Immunol. Microbial. 17,121-127.
28.
Midgley, Jr., A.R. (1966) Endocrinology
29.
Shaha, C., Suri, A., and Talwar, G.P. (1990) lnternat
30.
Goldberg, E. and Shelton, J.A. (1985) in lmmurwlogical ception
and Fertility,
79,10-18. J. Androl.
13,17-25.
Approaches
to Contra-
G.P. Talwar, Eds, PlenumPress,New York, 219-230.
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