Direct Antisperm Antibody Examination of Infertile Men

Transcription

1 Global Jounal of Medical eseach Volume 12 Issue 3 Vesion 1.0 May 2012 Tye: Double Blind Pee Reviewed Intenational Reseach Jounal Publishe: Global Jounals Inc. (USA) Online ISSN: & Pint ISSN : Diect Antisem Antibody Examination of Infetile Men By Saad S. Al-Dujaily, Wathic K. Chaki & Sundus F. Hantoosh Univesity of Al-Nahain/Iaq Abstact - The study was conducted to elucidate the coelation between diect antisem antibody (ASA) test and some semen chaactes of infetile men at the Institute of Embyo Reseaches and Infetility Teatment at Al-Nahain Univesity fom Mach 2009 to June Micoscoic and macoscoic semen analysis, Diect Sem MAR test and Diect Sem MAR test wee efomed on semen samles. Foty-two men wee included in the study, of whom, all undewent diect sem MAR test, but due to obseve bias, only 36 Diect sem MAR tests wee included in the study. Thee was a significant coelation between susected and viscosity (=0.65, = ); susected and H (=0.42, = ); and susected and liquefaction time (=0.44, =). A significant negative coelation was found between susected and ogessive sem motility (=-0.44, =); susected and susected with mohologically nomal sem ecentage (=-0.65, =0.03 and =-0.43, =, esectively); susected and sem agglutination (=- 0.54, =0.03). Keywods : Diect antisem antibody,,, immunological facto infetility GJMR -D Classification : FOR Code: , , Diect Antisem Antibody Examination of Infetile Men Stictly as e the comliance and egulations of: 2012 Saad S. Al-Dujaily, Wathic K. Chaki & Sundus F. Hantoosh. This is a eseach/eview ae, distibuted unde the tems of the Ceative Commons Attibution-Noncommecial 3.0 Unoted License htt://ceativecommons.og/licenses/by-nc/3.0/), emitting all non-commecial use, distibution, and eoduction inany medium, ovided the oiginal wok is oely cited.

2 Diect Antisem Antibody Examination of Infetile Men Saad S. Al-Dujaily α, Wathic K. Chaki σ & Sundus F. Hantoosh σ Abstact - The study was conducted to elucidate the coelation between diect antisem antibody (ASA) test and some semen chaactes of infetile men at the Institute of Embyo Reseaches and Infetility Teatment at Al-Nahain Univesity fom Mach 2009 to June Micoscoic and macoscoic semen analysis, Diect Sem MAR test and Diect Sem MAR test wee efomed on semen samles. Foty-two men wee included in the study, of whom, all undewent diect sem MAR test, but due to obseve bias, only 36 Diect sem MAR tests wee included in the study. Thee was a significant coelation between susected and viscosity (=0.65, = ); susected and H (=0.42, = ); and susected and liquefaction time (=0.44, =). A significant negative coelation was found between susected and ogessive sem motility (=- 0.44, =); susected and susected with mohologically nomal sem ecentage (=-0.65, =0.03 and =-0.43, =, esectively); susected and sem agglutination (=- 0.54, =0.03). This study, ecommends the use of Diect tests instead of the indiect assays in andology laboatoies in Iaq. Keywods : Diect antisem antibody,,, immunological facto infetility. I. INTRODUCTION I nfetility is a woldwide oblem which affects aoximately % of all coules, and is defined as, inability to conceive afte twelve months of contacetive-fee unotected intecouse (Ivine, 1998). Half of all the infetile coules emain unteated and/o unesolved. The main diagnostic gous fo infetility ae divided into those due to the male (25%), ovulation (25%), tubal, (20%), unexlained cause (25%) and endometiosis (5%) (Temleton, 1995).Among infetile coules, 40% ae imay due to the infetility of the male atne, while in 20% of these cases, it is a combination of both male and female factos (Agawal and Kulkani, 2003).The common causes of male infetility ae vaicocele, accessoy gland infection, immunological factos, congenital abnomalities, obstuctive azoosemia; iatogenic systemic and endocine causes. Ivine, (1998) eoted that the occuence of sontaneous antisem antibodies (ASA) to human Autho α : Deatment of Clinical Reoductive Physiology, IVF Institute, Univesity of Al-Nahain/Iaq. aldujaily8@yahoo.com Autho σ : Consultant Clinic, IVF Institute, Univesity of Al- Nahain/Baghdad/Iaq. sematozoa fom infetile men was fist descibed by Rumke and indeendently by Wilson in Antisem antibodies (ASA) ae the main cause of immunological infetility as they imai sem function by binding to the sem membane (de Keste and Bake, 1999). ASA can be detected in seminal lasma, whee they may be bound to the sem suface o solubilized in the seminal lasma, o in cevical mucus, oviductal fluid o follicula fluid of women. Natually occuing ASA in men can affect fetility by vaious mechanisms (Gadini et al., 1995). Some of them ae mainly elated to the extent of the sem autoimmunization (e.g. sem agglutination and imaied cevical mucus enetation); othes ae elated to immunoglobulin (Ig)-isotye (e.g. comlement-mediated sem injuy though the female genital tact); o to antigenic secificity of antisem antibodies (e.g. intefeence with gametes inteaction)( Gadini et al., 1995). Antisem antibodies can be defined as immunoglobulins of the,, and/o IgM isotye that ae diected against vaious asects of the sematozoa (head, tail, midiece o combination of them) (Temleton, 1995). Immunoglobulin subclasses and have been demonstated in the ejaculates of men with antisem autoimmunity. Though IgM seems to have no clinical imact, it is aely detected alone o combined with o (Hijot, 1999). In Iaq, due to difficulties and high exenses in imoting the kit of diect ASA test, the ocedues that diagnose the antisem antibodies in the andology laboatoies ae indiect assays, using sea of men. It has been eoted that anti-sem can be tested in seum but this is of little benefit as seum anti-sem neithe coelates with anti-sem Ig in semen, no influences fetility ognosis (Esteves et al., 2007; Agostini and, Lucas, 2011). Globally, semen samle assays of men ae used. Antibodies can be measued in cevical mucus and seminal lasma as well as in seum o as an antigen-antibody comlex on the suface of sem. Diect assessment of sem antibodies on the sem suface is efeed to indiect measuement because caacitation of sem might be useful in defining antibody eactivity (Esteves et al., 2007).This study is designed to find out the coelation between diect ASA test and cetain semen chaactes of infetile men. 37 Global Jounal of Medical Reseach Volume XII Issue III Vesion I 2012 Global Jounals Inc. (US)

3 Diect Antisem Antibody Examination of Infetile Men 38 2 Global Jounal of Medical Reseach Volume XII Issue III Vesion I II. METHODS Infetile males attending Institute of Embyo Reseaches and Infetility Teatment at AL-Nahain Univesity duing the eiod fom Mach 2009 to June 2010 wee included in this study. Seminal fluids wee collected by mastubation afte thee to five days of abstinence and seminal fluid analyses wee efomed within one hou of samle collection. Seminal fluid analysis was efomed accoding to WHO guide line (1999). Fo examle, sem concentation > 20 million/ml, ogessive sem motility (a+b=50%), mohologically nomal sem (> 30%), and ound cells (<5 cells e high owe field). All of the samles wee subjected to Diect sem MAR, and sem MAR tests (Fetio N.V. industieak Nood Beemem, Belgium). Diect Sem MAR and tests ae qualitative latex tests fo detection of sem antibodies of the and classes, esectively. Fo Diect Sem MAR test, the eagent used was Sem MAR latex aticles and fo Diect Sem MAR test, the eagents used wee Sem MAR latex aticles and Sem MAR antiseum. Fo both tests, on a micoslide 10 micolites of fesh semen and 10 micolites of Sem MAR latex aticles wee added. The samle and the latex eagent wee mixed five times with the edge of a cove glass. The cove glass was ut on the mixtue and the mixtue was obseved unde a light micoscoe at owe 40x magnification. The esult was ead afte 2-3 minutes. The latex aticles attached to motile sem wee obseved. One hunded sematozoa wee counted to detemine the ecentage of eactive sems. If no attachment of beads to sem was obseved, the eading was again efomed afte ten minutes as descibed in the infomation booklet accomaniny with the kits (Jage et al., 1978 ; Hinting, 1988). Samles with less than 10% eactivity wee labeled as "Nomal". The diagnosis of immunological infetility was "Susected" when 10-39% of the motile sematozoa wee attached to latex aticles. If 40% o moe of the sematozoa wee attached, immunological infetility was "ly Pobable" as mentioned in the instuctions accomanied with the kits (Jage et al., 1978). Based on these findings, the atients wee divided into thee gous fo each, and, into nomal, susected, o highly obable immunological infetility. III. STATISTICAL ANALYSIS The data was exessed as mean + standad eo (SE). Futhe analysis was accomlished using vicaiate Peason s coelation coefficient. P-value with less than 0.05 was consideed significant (Solie, 1995). IV. RESULTS Foty-two atients wee included in this study. Thei age anged between 22 to 45 yeas. Most of the atients had imay infetility (80.9%), wheeas the emaining atients had seconday infetility. Foty-two men wee subjected to Diect Sem MAR and Sem MAR tests. test fo thity-six out of the total foty-two samles wee included in the study; the emaining six wee excluded because of bias in thei esults. Table 1 shows no significant coelation between and with semen volume (=0.27, =0.22) and (=-0.18, =0.44) esectively. Howeve, thee was a significant negative coelation between nomal and semen volume (=-0.54, =0.03). A significant coelation was obseved between susected and viscosity (=0.65, =), and between susected and H (=0.42, =). Thee was a significant coelation between susected and liquefaction time (=0.44, =). Table 2 shows the coelation between both and with micoscoic semen analysis. A significant coelation was noticed between high and sem concentation(=0.46 =) with a significant negative coelation between susected and ogessive motility (-0.44, =). Thee was also a significant coelation between susected and susected with mohologically nomal sem ecentage (=-0.65, =0.03 and =-0.43, =, esectively). The coelation coefficient fo susected and sem agglutination had a significant negative value (=- 0.54, =0.03). V. DISCUSSION The esults evealed no coelations of both and with semen volume, liquefaction time and H, which ae all nomal in immunological infetility. Howeve, thee was a negative coelation between nomal and semen volume. This can be inteeted by the fact that in a samle with a low semen volume, nomal level is an indicato of accessoy glands disode athe than an immunological facto as the undelying cause of infetility. Seveal authos have stated that thee is no elation between nomal antibodies value and abnomal semen aametes (Soffe et al., 1990). The othe coefficient coelation was found between susected and liquefaction time, which means that fo each semen samle taking a longe time fo liquefaction, moe eactivity could be detected. A ositive coelation of susected with viscosity and H was also obseved. This finding may be due to acute o chonic genitouinay tact infections, asymtomatic Chlamydia tachomatis infections, o an infection with human immunodeficiency vius (Naz et al., 1990; Soffe et al., 1990 ).It is thought that locally 2012 Global Jounals Inc. (US)

4 Diect Antisem Antibody Examination of Infetile Men oduced is the most fequently imlicated of humoal immune infetility (Fancavilla et al., 2007). In this study, we noticed a negative coelation between susected and sem motility, sem mohology and sem agglutination.these coelations emhasized the cause of infetility. Antisem antibodies that ae located on the tail of sem can cause the sem to become immobilized o clumed togethe; and when antisem antibodies ae found on the head of sem, they can event the sem fom being able to efficiently make its way though a woman's cevical mucus to the ovum (Agostini and Lucas, 2011). A significant incease in the ootion of motile sems involved in agglutination has been eoted in the esence of antisem antibodies (Hijot, 1999). Most of the men with antisem antibodies had both, and on thei sematozoa but some had only. Pue eactions wee not ecoded and this is consistent with othe studies (Bohing et al., 2001). An osonizing effect is exeted by antisem antibodies which enhance sem hagocytosis and lysis by eitoneal macohages of the female and this effect is mediated by Fc-eceto fo (Agostini and Lucas, 2011). Cytotoxic effects due to comlement activation can be oduced by but not sem bound antibodies (Hinting, 1988). Abeation of the immune homeostasis may give ise to "immunological infetility". The majoity of subjects with asthenozoosemia and/o oligozoosemia have a highe incidence of low comlement-inhibiting activity and a educed level of comlement egulatoy oteins in thei seminal lasma, such that nonsecific activation of the altenative comlement athway occu, inflicting injuy to sem (Soffe et al.,1990). In this study, six infetile atients showed significant high ecentages of with no agglutination in thei semen. One exlanation is that the autoimmune disease is at its initial stages. If those atients emained unteated fo a cetain eiod of time and wee subsequently subjected to seminal fluid analysis test, notable agglutination might be noticed. Howeve, thee is vaiety of teatments available to hel the coules stuggling with ASA-mediated infetility, such as coticosteoids, inta-uteine insemination by sem washing and in-vito fetilization ogams (Fancavilla et al., 1997). Diect sem MAR and Diect sem Ma tests ae quick, easy, and sensitive, but equie motile sematozoa to ensue that the eythocytes o immunobeads adhee to the sem membane (Jage et al., 1978). In seveal counties, Diect sem Ma test kits ae used to detect antisem- in semen. Positive and dubious samles ae subsequently tested fo antisem. antibodies ae moe significant clinically, but vey aely occu without associated ; theefoe, the test of antisem antibodies in semen is sufficient as the initial sceening ocedue (Naz et al., 1990). Diect sem MAR test is commecially available, chea and quicke, and ae geneally consideed as moe suitable fo outine sceening of all semen analyses (Fancavilla et al.,1997). Diect and indiect tests which do not detect the egional secificity of antibody link o which lack secificity fo suface antigens because of concomitant exosue of intenal antigens, ae not suitable fo delineating a diagnosis (Hinting et al., 1988). One of the most ational ways to test fo antisem antibodies in males is by means of the diect mixed agglutination eaction (MAR) test (Bohing et al., 2001). This test is quick, easy, and sensitive, but equies motile sematozoa to ensue that the eythocytes o immunobeads adhee to the sem membane. It is an excellent sceening test if the eaction is negative o weakly ositive (Bohing et al., 2001). In addition, sem MAR is commecially available and equies no sem ocessing (Esteves et al., 2007). REFERENCES RÉFÉRENCES REFERENCIAS 1. Agawal, H.S., and Kulkani, K.S Efficacy and safety of seman in atient with oligosemia : an oen clinical study. Indian J. Cl. Pactice. 14 (2): Agostini, A., and Lucas, H Semen analysis. 9th Postgaduate Couse fo Taining in Reoductive Medicine and Reoductive Biology, Geneva, Oct. 3. Bohing, C., Kause, E., Habemann, B., and Kause, W Isolation and identification of sem membane antigens ecognized by antisem antibodies, and thei ossible ole in immunological infetility disease. Mol. Hum. Reod. 7(2): de Keste, D.M., and Bake, H.W Infetility in men. ecent advances and continuing contovesies. J. Clin. Endocinol. Metab. 84(10): Esteves, S., Schneide, D., and Veza, S Influence of antisem antibodies in the semen on intacytolasmic sem injection outcome. Intenational Baz. J. Uol. 33 (6): Fancavilla, F., Romany, R., Santucci, R., Maone, V., Poezi, G., Ruvolo, G Intefeence of antisem antibodies with the induction of the acosome eaction by zone ellucida (ZP) and its elationshi with the inhibition of ZP binding. Fetil. Steil. 67(6): Fancavilla, F., Santucci, R., Babonetti, A., and Fancavilla, S Natually-occuing antisem antibodies in men: intefeence with fetility and clinical imlications.an udate. In Font Biosci. 12: 39 Global Jounal of Medical Reseach Volume XII Issue III Vesion I 2012 Global Jounals Inc. (US)

5 Diect Antisem Antibody Examination of Infetile Men Gadini, L., Lenzi, A., Culasso, F., Lombado, F., Paoli, D., Dondeo, F Study of antisem antibodies bound to the sem cell suface and thei elationshi to ciculating ASA. Am. J. Reod. Immunol. 34(6): Hijot, T Antisem antibodies: Antisem antibodies and infetility: an unsolvable question? 10. Hinting, Hum. Reod. A., Vemeulenl, 14 (10): L. and Comhaie, F The indiect mixed antiglobulin eaction test using a commecially available kit fo the detection of antisem antibodies in seum. Feti. Steil. 49(6): Ivine, D.S Eidemiology and aetiology of male infetility. Hum Reod. 13, (sul 1)1: Jage, S., Keme, J., van Slochteen-Daaisma, T A simle method of sceening fo antisem antibodies in the human male. Detection of sematozoal suface with the diect mixed antiglobulin eaction caied out on unteated fesh human semen. Int. J. Fetil. 23(1): Naz, R.K., Ellauie, M., Phillis, T.M., Hall, J Antisem antibodies in human immunodeficiency vius infection: effects on fetilization and embyonic develoment. Biol Reod. 42(5-6): Soffe, Y., Ron-El, R., Golan, A., Heman, A., Casi, E., Sama, Z Male genital mycolasmas and Chlamydia tachomatis cultue: its elationshi with accessoy gland function, sem quality, and autoimmunity. Fetil. Steil. 53(2): Solie, D.E Medical biostatistics: Examination and boad eview. Nowalk, Connecticut: Aleton and Lang; : Temleton, A Infetility-eidemiology, aetiology and effective management. Health Bull. 53 (5): Global Jounal of Medical Reseach Volume XII Issue III Vesion I Kind of Ig(n) Nomal Susected 6 Nomal 18 Susected 21 3 Table 1 : Coelation of and with macoscoic seminal fluid aametes. Mean+SE In % 5.47± ± ± ± ± ±0.24 Values ae Mean+SE = Coelation coefficient = Pobability Volume In ml 2.92± ± ± 2.88± ± ± * Liquefaction Time 36.0± ± ± 38.65± ± ± * Viscosity 0.20± ± 0.2± 0.38± 0.38± 0.38± * H 7.76±- 7.76± 7.76± ± ± ± * Global Jounals Inc. (US)

6 Diect Antisem Antibody Examination of Infetile Men Table 2 : Coelation of and with micoscoic seminal fluid aametes. Kind of Ig Reactivi ty (n) Nomal Susect ed 6 Nomal 18 Susect ed 21 3 Mean + SE 5.47±0.06 Count 52.05± ± ± ± ± ± ± ± ± ± ±1.21 Values ae Mean ± SE = Coelation coefficient = Pobability * Pogessive motility 39.2± ± ± ± ± ± * \ WBC+ gem cells 1.2± ± ± ± ± ± Mohology ± ± ± ± ± ± * * Agglutinati on 5.25± ± ± ± ± ± * Round Cells 8..35± ± ± ± ± ± * Global Jounal of Medical Reseach Volume XII Issue III Vesion I 2012 Global Jounals Inc. (US)

7 Diect Antisem Antibody Examination of Infetile Men 42 2 Global Jounal of Medical Reseach Volume XII Issue III Vesion I This age is intentionally left blank 2012 Global Jounals Inc. (US)