Recurrent miscarriages, innate immunity, and autoimmune reaction to chlamydial 60-kDa heat shock protein--is there an association?

OBJECTIVE: To evaluate a potential association of immunity to the Chlamydia trachomatis 60kDa heat shock protein (ChlamHSP60) and recurrent miscarriages. DESIGN: Prospective study. SETTING: Outpatient miscarriage clinic of a university-based hospital. PATIENT(S): 120 asymptomatic women with a history of recurrent miscarriages. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Determination of serum immunoglobulin G (IgG) antibodies (Ab) to ChlamHSP60 and human HSP60 and, in parallel, mannose-binding lectin (MBL) and the total hemolytic complement (CH50); medical history and clinical examination, including multiple relevant laboratory determinants. RESULT(S): ChlamHSP60 Ab were detected in 24 (20%) of 120 patients. Antibodies to human HSP60 were found in 19 (15.8%) of 120 patients, and more frequently in individuals who tested positive for ChlamHSP60. ChlamHSP60 were statistically significantly associated with antichlamydial IgG Ab. However, antibodies to ChlamHSP60 were not related to medical history, the number of abortions, or the time frame of fetal loss. ChlamHSP60 antibodies were not associated with the relevant variables of the coagulation cascade, a panel of autoimmune parameters including thyroid autoimmunity, deficiencies of the complement system (low MBL), or with antibodies to common infectious diseases. No statistically significant differences were was found when comparing the prevalence of ChlamHSP60 Ab in the study group with recurrent miscarriages and 90 controls (women attending for an annual pelvic examination). CONCLUSION(S): Immunity to ChlamHSP60 does not play a major role in the etiology of recurrent miscarriages.

Medical history screening for thrombophilic risk: is this adequate?

OBJECTIVE: To evaluate the reliability of medical history taken before hormonal medication administration to identify women with an increased risk for thromboembolic events detected by laboratory screening. DESIGN: Prospective study. SETTING: Outpatient endocrine clinic of a university-based hospital. PATIENT(S): Four hundred forty-three consecutive women (median age 49 years) who presented with endocrine disorders. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Parallel screening, on first visit, with a complete medical history, and same-day laboratory screening for thromboembolic risk. Laboratory examination in a two-step procedure with a standard assay and confirmation by genotyping on the second visit. RESULT(S): A total of 13.8% (61/443) patients with an abnormal activated protein C (APC) resistance test were identified. Second blood samples revealed a prevalence of factor V (Leiden) heterozygosity in 10.9% (homozygosity in 0.2%). There was a significantly higher prevalence of thrombotic events in a first degree relative of patients with APC resistance (in 18.3%) compared with women with a normal test outcome (in 7.8%). However, medical history (personal and family history) was negative concerning hints for thromboembolic events in more than 80% of patients with a laboratory risk profile for thromboembolic morbidity. No association of APC and factor V testing with the patients' gynecological/obstetric history (e.g., live birth rate, miscarriages) was observed. CONCLUSION(S): Medical history alone may be inadequate to identify all patients at risk for thromboembolic complications with hormonal treatment.

Are chlamydial lipopolysaccharide-directed antibodies in seminal plasma or serum clinically significant during investigation of male infertility?

OBJECTIVES: To analyze the clinical relevance of chlamydial serology in seminal plasma (SP)/serum for male fertility. METHODS: We determined the IgG, IgA, and IgM class antibodies to Chlamydia trachomatis with a lipopolysaccharide-directed standard enzyme-linked immunosorbent assay in the serum of 173 asymptomatic subfertile couples and in the same-day SP in a prospective study. The semen quality evaluation was done using aliquots of the same ejaculate (eg, sperm analysis, leukocytes, bacterial cultures, sperm functional capacity). The medical history, clinical examination, postcoital testing, and determination of subsequent fertility were also done. RESULTS: The antichlamydial antibodies of the 3 classes were significantly interrelated, and SP Chlamydia IgA antibodies were significantly associated with the findings in the serum. The evidence for previous or persistent chlamydial infection was significantly more frequent in the female partners of men with Chlamydia antibodies in the serum and/or Chlamydia IgA antibodies in the SP. The chlamydial serology results in the SP/serum were not associated with the sperm quality, leukocytes, semen culture findings, or sperm functional capacity. CONCLUSIONS: The chlamydial serology results (ie, IgG, IgA, and IgM antibodies in SP/serum) determined using a lipopolysaccharide-directed enzyme-linked immunosorbent assay were not indicative of reduced sperm function or subsequent fertilizing capacity.

Cytomegalovirus (CMV) infection--related to male and/or female infertility factors?

OBJECTIVE: To screen for cytomegalovirus (CMV) infection in male and female partners of subfertile couples and to analyze a potential relationship with clinically relevant parameters of male and female factor fertility. DESIGN: Prospective study. SETTING: Outpatient infertility clinic of a university-based hospital. PATIENT(S): Randomly selected male and female partners of asymptomatic subfertile couples. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Screening for CMV in the semen of the male partner (using nested polymerase chain reaction) and, at the same time, in the endocervical material of the female partner; same-day CMV serology (anti-CMV IgG and IgM class antibodies [Ab]); medical history and clinical examination; evaluation of semen quality, including sperm functional capacity and detection of antisperm Ab (ASA) and seminal white blood cells (WBC) in aliquots of the same ejaculates; bacterial screening of both partners; and detailed examination of the cervical factor and other variables of female subfertility. RESULT(S): The presence of CMV in semen was not significantly related to semen quality including sperm functional capacity, local antisperm Ab, or seminal WBC. CMV in endocervical material was not associated with a reduced quality of the cervical mucus or with other female infertility factors. CMV in semen was not associated with the presence of CMV in the endocervical material of the female partners. CMV infection was not significantly associated with other microorganisms of the lower genital tract. CMV serology (IgG and/or IgM Ab) did not sufficiently reflect CMV presence in semen or the cervix. CONCLUSION(S): CMV presence in the genital tract of subfertile patients is considerable, but findings do not suggest that sexual transmission is a frequent route of infection or that CMV infection is a significant cause of infertility.

Role for tumor necrosis factor alpha (TNF-alpha) and interleukin 1-beta (IL-1beta) determination in seminal plasma during infertility investigation.

OBJECTIVE: To evaluate the clinical relevance of tumor necrosis factor alpha (TNF-alpha) and interleukin 1-beta (IL-1beta) determination in seminal plasma during infertility investigation. DESIGN: Prospective study. SETTING: Outpatient infertility clinic of a university-based hospital. PATIENT(S): Randomly chosen asymptomatic males (n = 148) from subfertile couples. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Determination of TNF-alpha and IL-1beta in seminal plasma (SP) by enzyme-linked immunosorbent assay (ELISA). In aliquots of the same ejaculates: 1) evaluation of semen quality with sperm analysis and sperm function testing; 2) determination of antisperm antibodies (ASA) of the immunoglobulin (Ig) G and IgA class; 3) microbial screening; and 4) immunocytochemical round cell differentiation to determine leukocyte counts and ratios. Medical history, clinical examination, and determination of subsequent fertility (after control for female infertility factors). RESULT(S): The concentrations of TNF-alpha and IL-1beta in SP correlated significantly (r = 0.65; P<.0001), and these parameters were significantly related to the leukocyte ratio (%LC) of the seminal round cells (r = 0.36; P<.001) and the leukocyte counts per ejaculate (r = 0.34; P<.001). There was no relationship of TNF-alpha and IL-1beta levels in SP with semen quality or parameters of sperm functional capacity, and there was no association with local ASA of the IgG or IgA class. The concentration of both cytokines was also not related to the outcome of the microbial screening and did not affect subsequent fertility. No correlation of TNF-alpha and IL-1beta levels in SP with the concentration of C-reactive protein in same-day serum samples was found. CONCLUSION(S): The levels of TNF-alpha and IL-1beta in seminal fluid correlate significantly with leukocyte counts and ratios in the same ejaculates, as indicators of silent male genital tract infection/inflammation. However, this is not related to semen cultures in asymptomatic individuals and not associated with clinically relevant parameters of semen quality, including sperm fertilizing capacity.

High resolution proton magnetic resonance spectroscopy of human cervical mucus.

High resolution 1H NMR spectroscopy is a powerful method for qualitative and quantitative analysis of the highly viscous human cervical mucus (CM). Up to 23 compounds could be identified in this study and can be observed in the 1H NMR spectra of native mucus without the need of any complicated preparative chemistry. Storage conditions could be excluded as a possible reason for variations observed between different samples. pH values decreased after freezing and storing at 253 K. For NMR studies, lyophilization proved to be most useful, allowing the determination of the water content, replacement of H2O by D2O, and most importantly, absolute quantification of low molecular mass compounds. In a small collective of women the concentrations of some of the small constituents of the CM are strongly correlated; an example is the mutual positive correlation of taurine, citrate and creatinine. In conclusion, high resolution 1H NMR spectroscopy is a valid method to investigate mucus composition and to determine absolute concentrations of low molecular mass compounds in CM.

Seminal antibodies to human 60kd heat shock protein (HSP 60) in male partners of subfertile couples.

BACKGROUND: Heat shock proteins (HSP) are essential mammalian and bacterial stress proteins. At the cellular level, they act as chaperones, have important regulatory functions, and are considered to be an essential factor for reproduction. Scarce information exists on the role of sensitization to HSP and the potential role in the aetiology of male infertility. METHODS: The potential association of immunoglobulin (Ig)A antibodies (Ab) to the human 60 kDa heat shock protein (HSP 60) with several parameters of subclinical male genital tract infection/inflammation and with semen quality and sperm fertilizing capacity was analysed in a prospective study. IgA Ab to human HSP 60 were determined in seminal plasma of 202 randomly chosen male partners of subfertile couples with a median duration of infertility of 4 years (range 1-15 years), who were asymptomatic for genital tract infection. After medical history and clinical examination, a comprehensive evaluation of semen quality, in aliquots of the same ejaculates used for HSP Ab determination, included: sperm analysis; local antisperm antibody (ASA) screening; standardized sperm-cervical mucus (CM) penetration testing; immunocytochemical round cell differentiation to determine seminal leukocyte counts; evaluation of complement fraction C(3) and of some pro-inflammatory cytokines; and microbial screening. Subsequent fertility was recorded after 6 months. RESULTS: The presence of HSP 60 IgA Ab in seminal fluid (total positive 6.9%) was significantly associated with leukocytospermia, the presence of C(3), and also with high interleukin (IL) levels in seminal plasma. HSP 60 Ab were not related to the bacterial colonization of ejaculates. There was no association of seminal IgA Ab to human HSP 60 with semen quality, determined with microscopical semen analysis, nor with local IgG- or IgA-class ASA. There was no relationship with sperm intrinsic motility and duration of motility in the sperm CM-penetration test, nor with sperm fertilizing capacity. CONCLUSIONS: The combined presence of IgA Ab to human 60 kDa HSP, leukocytes and other established infection/inflammation markers in semen might suggest a potential role of the immune response to heat shock proteins (HSP) in cases of silent male genital tract infection, but the results do not indicate a marked relationship of HSP 60 Ab in seminal fluid with standard parameters of semen quality.

Are zinc levels in seminal plasma associated with seminal leukocytes and other determinants of semen quality?

OBJECTIVE: To evaluate a potential association of zinc levels with seminal leukocytes, the outcome of semen cultures; and semen quality and sperm fertilizing capacity. DESIGN: Prospective study. SETTING: Outpatient infertility clinic of a university hospital. PATIENT(S): Two hundred fifty-six randomly chosen asymptomatic males from subfertile couples. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Determination of zinc in seminal plasma by flame atomic absorption spectroscopy. In aliquots of the same ejaculates the following tests were performed: immunocytochemical round cell differentiation to determine leukocyte counts and ratios, microbial screening, and comprehensive evaluation of semen quality (sperm analysis, biochemical parameters, antisperm antibody testing, and in vitro examination of sperm ability to penetrate cervical mucus). The patients underwent medical history, clinical examination, and postcoital testing. Subsequent fertility was determined (controlled for female infertility factors). RESULT(S): The concentration of zinc in seminal plasma did not correlate in a statistically significant way with leukocytes in semen, nor was it associated with bacterial colonization. There was no statistically significant relationship of zinc in seminal plasma or serum with semen quality parameters nor with local antisperm antibody testing of the IgG or IgA class. Zinc levels did not influence sperm capacity to penetrate cervical mucus in vitro or in vivo, and did not affect subsequent fertility. CONCLUSION(S): The zinc level in seminal fluid and serum is not associated with silent male genital tract infection (indicated by seminal leukocytes); nor is it related to semen cultures in asymptomatic individuals. The lack of association with other semen quality parameters indicates that the routine determination of zinc levels during infertility investigation is not recommended.