New insights into immunological infertility testing for anti-sperm antibodies and sperm-zona pellucida binding ability by the immunobead assay and sperm-hyaluronic acid interaction

The hyaluronic acid (HA)-rich extracellular matrix (ECM) of the cumulus oophorus is known to facilitate fertilisation. It has been suggested that HA may enhance fertilisation in a number of species, and in macaque sperm, HA has been shown to increase the number of acrosome reactions that follow sperm binding to the zona pellucida. In this study, we investigated the effects of HA on intracellular Ca2+ in capacitated cynomolgus macaque sperm. Fluorometry studies using the intracellular Ca2+ indicator Fluo-3 showed that addition of 100 μg/ml of HA induced a rapid increase in intracellular Ca2+. This Ca2+ increase (approximately 2–3 times above basal levels) was inhibited by preincubation of sperm with Fab fragments of anti-recombinant PH-20 IgG. The frequency of acrosome reactions in sperm exposed to HA was not above control levels. A synthetic gel was prepared with similar viscosity to the cumulus and with HA trapped in its matrix. Video imaging of individual sperm was used to demonstrate that capacitated sperm swimming into the HA gel had increased intracellular Ca2+ levels. Preincubation of sperm with Fab fragments of anti-PH-20 IgG inhibited the increased intracellular Ca2+ levels induced by the HA gel. Sperm in control gel (no HA) did not show increased intracellular Ca2+, while sperm in gel containing anti-PH-20 IgG showed increased Ca2+ (positive control). Sperm loaded with Fluo-3 were allowed to interact with cynomologus macaque cumulus masses, and sperm within the cumulus ECM clearly showed increased intracellular Ca2+ that was inhibited when sperm were preincubated in anti-PH-20 Fab. Fluorescein isothiocyanate (FITC)-HA was found to bind to sperm over the acrosomal region, corresponding to PH-20 localisation, and this binding could be inhibited by preincubation of sperm with anti-PH-20 fragments. The results of this study show that HA increases intracellular Ca2+ in macaque sperm through interaction with plasma membrane PH-20. We propose that HA binding to plasma membrane PH-20 induces an aggregation of receptors that in turn results in intracellular signalling. As a result, sperm have higher basal CA2+ levels and are more responsive to induction of the acrosome reaction after binding to the zona pellucida.

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Relationship Between Post-Pubertal Mumps Infection In Males With Infertilityand Its Effect on The Result of Seminal Fluid Analysis and Occurrence of Immunological Infertility

Diyala Journal of Medicine ◽

10.26505/djm.19025500814 ◽

2020 ◽

Vol 19 (2) ◽

pp. 118-128

Author(s):

Sahar Mohammed Zaki Abdullah ◽

Keyword(s):

Seminal Fluid ◽

Testicular Atrophy ◽

Enzyme Linked Immunosorbent Assay ◽

Adult Males ◽

Cross Sectional ◽

Fluid Analysis ◽

Immunological Infertility ◽

History Of ◽

Sperm Antibodies ◽

Mumps Infection

Background: Mumps caused by paramyxovirus in the same group as parainfluenza and Newcastle disease virus, orchitis is the widely recognized complication in post-pubertal males. Approximately 50% of patients with orchitis have some degree of testicular atrophy, but sterility is rare. Objective: To demonstrate the effect of the post-pubertal mumps infection on the result of seminal fluid analysis and to know if infection by mumps after puberty can lead to immunological infertility and production of anti-sperm antibodies. Patients and Methods: Cross-sectional study in which 300 infertile males attending microbiology laboratory in Rizgary teaching hospital in Erbil city in Iraqi Kurdistan from November 2017- September 2018. Questionnaire was prepared for each male which included: name, age, occupation, history of mumps infection after puberty. Seminal fluid collected from each patient after 3 days of abstinence and analyzed according to WHO guidelines. Seminal fluid and serum obtained from each infertile male for the detection of anti-sperm antibodies by ELISA (enzyme-linked immunosorbent assay). Results: The mean age of the participant in the study was (32.805 year) with the maximum and minimum age was (50, 18) years respectively. The incidence of post-pubertal mumps was (13.3%) and the incidence of abnormal seminal fluid analysis was (10%) and the highest abnormal seminal fluid variables among infertile males with post-pubertal mumps was oligoasthenozoospermia (43%) and the anti-sperm antibodies in the semen of infertile males with post-pubertal mumps infection was (60%) while the frequency of anti-sperm antibodies in the serum in infertile males with a history of post-pubertal mumps was (40%) which was higher than those with no history of post-pubertal mumps with highly significant relation statistically. Conclusion: Abnormal seminal fluid analysis result may be due to post-pubertal mumps and it can lead to (Asthenozoospermia, Oligospermia, and Teratozoospermia). The highest percentage of abnormal seminal fluid variables was Oligoasthenozoospermia the occurrence of anti-sperm antibodies can be the sequence of post-pubertal mumps and by itself can lead to abnormality in the seminal fluid analysis. Mumps can lead to male infertility by affecting the seminal fluid parameters and also by the production of anti-sperm antibodies which can lead to subfertility in adult males. Keywords: Paramyxovirus, post-pubertal mumps, anti-sperm antibodies

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