Immunomodulatory Treatment for Infertile Men with Antisperm Antibodies

Ivan Bubanovic1  Stevo Najman2, Slobodan Kojic3

 1Department of Obstetrics and Gynecology – “Medica Centre” – Nis, Serbia and Montenegro

2Institute for Biology, University Medical School – Nis, Serbia and Montenegro

3Department of Obstetrics and Gynecology – Medical Center – Paracin, Serbia and Montenegro

Fertility and Sterility, 2004; 81(3):S7-31: pp. 20.

The results were presented on the

52nd annual meeting and postgraduate course of the Pacific Coast Reproductive Society.

April 28-May 2, 2004. Rancho Mirage • California, USA. ID:/Article No: 943

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ABSTRACT

One of the important categories of subfertility or infertility in humans may be immunological diseases mediated by antisperm antibodies (ASA). Blood sera from 35 infertile men of different age with ASA positive ELISA test were examined for the serum level of ASA before and after treatments with 1a,25-dihydroxy-Vitamin-D3 and Dexamethasone. We treated 18 infertile men with Vitamin-D3/Dexamethasone during 30 days, 9 infertile men with Dexamethasone only during 30 days and 8 infertile men received no treatment. All the patients showed poor parameters of spermogram and high level of ASA serum concentration (>75 U/ml). Serum concentration of ASA in non-treated group (313 U/ml), Dexamethasone only treated group (288) and Vitamin-D3/Dexamethasone treated group (124 U/ml) were significantly different. Serum level of ASA in Vitamin-D3/Dexamethasone treated group was significantly lower compared to the level before the treatment (P<0.01). These findings could be explained by immunosuppressive and immunomodulatory effects of Dexamethasone and Vitamin D3 treatment.

Key Words: antisperm antibody, dexamethasone, infertility, vitamin D3.

INTRODUCTION

Autoimmunity to sperm may occur because sperm cell antigens are first expressed during sexual maturation, long after the prenatal period when immunological self-tolerance is induced [1,2]. Protection against autoimmunity is provided by the blood-testis barrier composed predominantly of Sertoli cells isolating the tubular content from the vasculature and limited lymphatic drainage of the testis [3]. Several other immunoregulatory mechanisms also play a significant role in the prevention of anti-sperm immunity, such as immunosuppressive factors of seminal plasma, as well as both systemic nonspecific and specific factors (immunoregulatory cells, cytokines, absence of co-stimulatory molecules expression etc.) [4]. In some cases, auto-immunization with sequestered sperm molecules happened after disruption of blood-testis barrier by disease and/or injuries [5,6]. Generally, humoral immune response such as ASA formation can be induced primarily during infectious and noninfectious inflammations, or by obstruction of testicular efferent duct [1,7]. The ASA was also induced after accidental and/or surgical injury of testicles, exposure to very low temperature or cryptorchism [5,6]. Subsequently, infertility can result from antibodies directly binding the sperm, or from aspermatogenesis due to allergic orchitis. A similar phenomenon occurs in vasectomized laboratory rodents and man [5]. Most affected individuals develop epididymal sperm granulomas and testicular degeneration associated with the formation of antisperm antibodies [5,8]. Finally, the presence of ASA reacting with antigens on the sperm considered typical and specific immunological infertility [8].

ASA can impair fertilization by several mechanisms. They can interfere with sperm motility by immobilizing or agglutinating the sperm, or interfere with sperm-cervical mucus interaction and disturb sperm transport [9]. ASA mediated impairment of fertilization can occur as interference of the penetration into the oocyte, and perhaps affect zygote development by impairing early cleavage, or even damaging the implantation process [10]. Whether ASA are involved in pregnancy loss is still debatable as no conclusive evidence is available, so that this subject needs further research [9,10].

Corticosteroids and Vitamin D3 can have profound effects on immune response in mammals on different levels and IgG responses to different antigens [11,12]. Therefore, we speculate that Vitamin-D3 and Dexamethasone can be useful in treatment of infertile, ASA positive patients.

 MATERIALS AND METHODS

Thirty-five infertile men with serum level of ASA higher than 75 U/ml (as recommended by the ASA Kit manufacturer) and poor parameters of spermogram comprised three study groups.

Group 1

Eighteen men were treated with Vitamin D3/Dexamethasone combination. Vitamin D3 was administered orally (0.025 mcg/kg of body weight) during 30 days. Dexamethasone was administered during 30 days in dose-decreasing manner. On day one of the treatment, Dexamethasone was administered intramuscularly (im.) in one-day dose 110/mcg/kg. This was followed by one-day dose of 55 mcg/kg administered im. (days 2 and 3). The treatment with Dexamethasone was followed by decreasing oral dose starting with 42 mcg/kg (on day 4 of the treatment) to 7 mcg/kg (on day 30 of the treatment).

Group 2

Nine men treated only with Dexamethasone that administrated during 30 days as well as in group 1.

Group 3

Eight men were not treated and they will be included in treatment in one of further investigations.

Spermogram

The semen analyses were performed according to the guidelines of the WHO (1992) [13]. Sperm count in all groups of patients was analyzed before the treatment and 15 days after treatment. Abstinence time before sperm sampling was 5 days.

Serum Antisperm Antibody ELISA Test

Serum concentration of antisperm antibodies was performed on HUMAN ELISA READER instrument with Immuno-Biological Laboratories (IBL) Sperm Antibody Enzyme Immunoassay Kit. The test is based on a non-competitive ELISA. The strips were incubated with diluted sera (1:50) from patients, and after washing steps, were incubated again with peroxidase conjugated anti-human-Ig (IgA, IgG and IgM). Following the final wash and enzyme substrate addition, the developed color was determined using the ELISA reader. Positive results are indicated by ASA concentrations > 75 U/ml in diluted sample of serum as recommended by IBL.

Statistical Analysis

All parameters of 3 study groups were analyzed. P value of less than 0.05 was considered to indicate statistical significance. Calculations were performed with MS Excel ® 2002 by t-test.

RESULTS

Average age of all patients was 35±7. As a possible etiologic factor of ASA presence, we found cryptorchism in 11% (unilateral and bilateral), orchitis in 11%, varicocoela in 26%, accidental trauma in 3%, surgical intervention in 3%, epididymitis in 11% and unknown etiology in 35% of patients. Basic parameters of spermogram and serum level of ASA studied at the time of starting therapy and after 45 days are summarized in table 1.

Table 1 demonstrates basic parameters of spermograms before and after treatment in all 3 groups of infertile men. No significant differences were found recorded in sperm count volume, sperm concentration and percent of cells with normal morphology before and after treatment in 3 studied groups (P>0.05). Likewise, no significant differences in motility and viability of spermatozoa were found before and after treatment in group 2 and group 3 (P>0.05). Percent of motile (P=0.021) and vital (P<0.01) spermatozoa in group 1 is significantly higher after treatment in relation to the percent before treatment.

In group 3, no significant changes were found in serum level of ASA in relation to the initial sample and after 45 days (P=0.54). Also, serum level of ASA in group 2 did not show significant changes before and after the treatment (P=0.21). Nevertheless, the level of ASA in group 1 was found to be significantly lower after Vitamin D3/Dexamethasone treatment as compared to the level before the treatment (P<0.01).

Table 1. Basic parameters of spermograms and serum level of ASA before and after treatments (45 days).

 

 

 

Group 1

Vitamin D3 and Dexamethasone

n=18

Group 2

Dexamethasone only

n=9

Group 3

no treatment

n=8

Volume (ml)

before

2.5±0.7

2.6±0.7

2.8±1

after

2.6±0.7

2.7±0.5

2.9±0.4

Sperm concentration (106/ml)

before

17±9

16±11

17±9

after

19±9

17±9

19±7

Motile (%)

(after 60 min.)

before

34±17

34±5

34±13

after

45±15

35±7

34±8

Vital (%)

(after 60 min.)

before

41±8

37±9

38±6

after

52±10

39±9

36±8

Normal

morphology (%)

before

61±15

64±11

63±6

after

62±12

62±13

61±8

Serum level of ASA (U/ml)

before

311±77