Serum Autoantibodies in Mice Immunized with Syngeneic Testicular Germ Cells Alone

Abstract

These results indicate that titers and kinds of autoantibodies in TGC-immunized mice increase with EAO development and they are specific to TGC and ES.

Low-dose exposure to di-(2-ethylhexyl) phthalate (DEHP) increases susceptibility to testicular autoimmunity in mice

“…ELlSA was performed according to the method described by Hirai et al [30]. To prepare the antigens, TGC from untreated 10-week-old mice (n = 6) were homogenized separately in a carbonate-bicarbonate buffer (Sigma Chester, PA, USA; 1:1 000).…”

Specific autoantigens identified by sera obtained from mice that are immunized with testicular germ cells alone

“…At 4 weeks of age, in addition to this band, a band of approximately 42 kDa was detected in TGC proteins. At 8 weeks of age, three TGC protein bands (approximately 20, 28, and 60 kDa) were also detected 32 . Considering the EAO-inducing factors and expression levels in testis at 2 weeks and 8 weeks, the EAO-related proteins were 24–35, 42, and 60–75 kDa.…”

“…In addition to these, bands of 24–35 kDa and 60–75 kDa were detected in testicular proteins at 8 weeks of age 47 . Moreover, we incubated immunoblots with EAO serum from various weeks after TGC immunization 32 . At 0 week of age, a band corresponding to approximately 40 kDa was detected in TGC proteins, indicating the presence of natural autoantibodies against TGC.…”

“…ELlSA was performed using the method described by Hirai et al . 32 . For preparation of antigens, TGC obtained from 10-week-old normal mice (n = 3) was homogenized in carbonate–bicarbonate buffer (Sigma-Aldrich, MO, USA).…”

“…We have previously established another EAO model induced in both A/J and C3H/He mice with a very high incidence by two subcutaneous injections of viable syngeneic TGC without using any adjuvant 21 . Our EAO model is unique because serum autoantibodies are only against acrosomal regions of sperm and spermatids, but not Sertoli cells, Leydig cells and seminiferous tubular basement membrane 3 25 26 32 . This model showed that the immunologically privileged state of the testis is easily overcome using only two TGC injections.…”