猪血清及某些血清免疫球蛋白对猪精子的作用

EFFECTS OF SERA AND SOME SERUM IMMUNOGLOBULINS ON SPERMATOZOA IN PIG

新鲜猪血清对猪精子具有毒性作用。灭活补体的血清失去了杀精子作用，但使精子产生头对头凝集。分子筛层析证明，血清凝精子活性存在于富含免疫球蛋白的洗脱峰内。从５头处女猪的混合血清中制备出电泳纯的ＩｇＭ和ＩｇＧ（ＩｇＧ２和ＩｇＧ２的混合物）、ＩｇＭ在０．０２５－０．０５ｍｇ／ｍｌ（３×１０７个精子）时就引起明显的精子凝集，而ＩｇＧ在浓度为１．４１ｍｇ／ｍｌ（３×１０７个精子）时仍无凝精子作用。精浆对血清引起的精子凝集有抑制作用。在自身血清中，被精浆的猪红细胞自动凝集。环状沉淀显示，由分子筛层析得到的具有凝精子活性的洗出液可与精浆作用产生沉淀。双向免疫扩散显示，精浆跟血清作用形成一条沉淀线。以上结果提示：新鲜血清的杀精子作用可能与补体反应有关；ＩｇＭ是血清中主要的精子凝集团子；精子凝集可以跟精浆中的某种物质有关，这种物质可以竞争性地抑制血清的凝精子作用。

Twelve fresh serum samples were made from five gilts, four sprayed gilts and three castrated boars. All the samples were lethal to spermatozoa of four boars (Large-Yorkshire, Hampshire,Beterland and Duroc). The spermatozoa which were suspended in 1 ml of serum in final density of 3 × 107 were all killed in an hour. Microscopic examination of stained sperms revealed that more than 98% of the acrosomes of spermatozoa killed with sera were destroyed. The spermicidal activity disapperaed after fresh sera being heated at 56℃ for 30 minutes. But the inactivated sera induced head-to-head agglutination of spermatozoa.The SAT(sperm agglutination test)titers of a mixed serum of five gilts in four boars were 1024, 1024, 512, 256 respectively. Spermagglutinating activity was found in the γ-globulin-rich peaks which were eluted from Sephadex G-200 fractionation of serum. IgM and IgG (a mixture of IgG1 and IgG2) were isolated from the five gilts' mixed serum by means of ammunium sulfate fractionation. DEAE-cellulose chromatography and Sephadex G-200 chromatography. The purified IgM induced agglutination at the concentration of 0. 05- 0. 025mg/ml(3 ×10~7sperm)while the IgG did not at concentrations up to 1. 41mg/ml(3 × 10~7sperm). The serum-induced agglutination of sperm was inhibited by seminal plasma. The SAT titers of the five gilts' mixed serum in four boars were reduced to 4-32 when 10%seminal plasma was included in the serum dilutions. When the red cells and serum were made from the same pig,the red cells coated with seminal plasma agglutinated in the serum. the serum fractions that induced agglutination of sperm were able to react with seminal plasma and produce precipitate. one precipitin band was observed when seminal plasma was subjected to Ouchterlongy double diffusion against pig serum. These results suggest: 1. Fresh pig sera are cytotoxic to boar spermatozoa, the serum spermicidal activity may be related to complement fixation. 2. Inactivated sera induce head- to-head agglutination of spermatozoa and IgM is a major spermagglutinin. 3. The serum- induced agglutination of sperm is related to a certain substance in seminal plasma which can bind to spermatozoa and inhibit agglutination competitively.