A Schistosoma mansoni 62-kDa band is identified as an irradiated vaccine T-cell antigen and characterized as calreticulin.

The Schistosoma mansoni soluble adult worm antigen (SAWA) bands of 62/60 kDa were found to contain immunodominant T-cell immunogen(s) in irradiated cercariae-immunized Swiss and C57BL/6 mice. In the present study, spleen T cells of BALB/c mice immunized twice with ultraviolet light-irradiated cercariae proliferated and produced interleukin 4 in response to the 62/60-kDa SAWA bands in T-cell western assays. To characterize the 62/60-kDa bands, an adult S. mansoni worm cDNA expression library constructed in lambdagt11 was immunoscreened with serum of mice immunized with the 62/60-kDa antigens, and the immunoreactive cDNA inserts were sequenced. Purified 62- and 60-kDa proteins were used for amino acid microsequencing and for immunization studies in Swiss, C57BL/6, and BALB/c mice and rabbits. Taken together, the data indicated that the 60-kDa molecules are poorly immunogenic in mice and rabbits, whereas the 62-kDa species identified as S. mansoni calreticulin, is a good T- and B-cell antigen and represents a potential vaccine candidate.

Intracytoplasmic sperm injection in obstructive and non-obstructive azoospermia.

We compared the results of intracytoplasmic sperm injection (ICSI) in: (i) obstructive versus non-obstructive azoospermia, (ii) obstructive azoospermia using epididymal versus testicular spermatozoa and (iii) acquired versus congenital obstructive azoospermia due to congenital absence of the vas deferens (CAVD). A retrospective analysis was done of 241 consecutive ICSI cycles done in 103 patients with non-obstructive azoospermia and 119 patients with obstructive azoospermia. In the obstructive group, 135 ICSI cycles were performed. Epididymal spermatozoa were used in 44 cycles and testicular spermatozoa in 91 cycles. In the non-obstructive group, 106 cycles were performed. The fertilization and pregnancy per cycle rates were 59.5 and 27.3% respectively using epididymal spermatozoa, 54.4 and 31.9% respectively using testicular spermatozoa in obstructive cases, and 39 and 11.3% respectively in non-obstructive cases. The fertilization and pregnancy per cycle rates were 56.6 and 37% respectively in acquired obstructive cases, and 55.2 and 20.4% respectively in CAVD. In conclusion, ICSI using spermatozoa from patients with acquired obstructive azoospermia resulted in significantly higher fertilization and pregnancy rates as compared to CAVD and non-obstructive cases.

Fertilization and pregnancy rates after intracytoplasmic sperm injection using ejaculate semen and surgically retrieved sperm.

OBJECTIVE: To compare the fertilization rates and pregnancy rates (PRs) in intracytoplasmic sperm injection (ICSI) using sperm from ejaculates of normal and abnormal semen, epididymal sperm, and testicular sperm of obstructive and nonobstructive azoospermic patients. DESIGN: Retrospective study. SETTING: The Egyptian IVF-ET Center. PATIENT(S): Three hundred fifty patients underwent 366 ICSI cycles. INTERVENTION(S): ICSI, epididymal sperm aspiration, and testicular biopsy. MAIN OUTCOME MEASURE(S): Fertilization rates and PRs. RESULT(S): Patients were divided into five groups according to the quality and source of sperm. Patients in group 1 underwent 102 cycles of ICSI using ejaculated abnormal semen, group 2 underwent 44 cycles using epididymal sperm, group 3 underwent 82 cycles using testicular sperm from obstructive azoospermia, group 4 underwent 80 cycles using testicular sperm from nonobstructive azoospermia, and group 5 underwent 58 cycles using normal semen. There was no significant difference in the fertilization rates and PRs among groups 1, 2, and 3. In group 4, the fertilization rate and PR were significantly lower than in all other groups. In group 5, the fertilization rate was significantly higher than in all other groups. CONCLUSION(S): The fertilizing ability of sperm in ICSI is highest with normal semen and lowest with sperm extracted from a testicular biopsy in nonobstructive azoospermia. There was no significant difference in fertilization rates and PRs between ejaculated sperm of different parameters and surgically retrieved sperm in obstructive azoospermia.

Intracytoplasmic sperm injection using surgically retrieved epididymal and testicular spermatozoa in cases of obstructive and non-obstructive azoospermia.

This was a retrospective study of 115 patients who underwent 124 cycles of ICSI using surgically retrieved spermatozoa. The objective was to compare the results of ICSI in patients with obstructive azoospermia using epididymal spermatozoa (36 cycles) or testicular spermatozoa (58 cycles) with ICSI in patients with non-obstructive azoospermia using testicular spermatozoa (30 cycles). When epididymal spermatozoa were used for ICSI, the fertilization rate per injected metaphase-II oocyte and the clinical pregnancy rate per ICSI cycle were 60.4 and 25%, respectively. When testicular spermatozoa were used in obstructive cases, the fertilization rate and pregnancy rate were 57.9 and 34.5%. In non-obstructive cases the fertilization and pregnancy rates were 41.2 and 16.6%. When patients with obstructive azoospermia were regrouped according to the cause of obstruction, the fertilization and pregnancy rates were 59.1 and 35.1% in acquired obstruction and 58.7 and 24.3% in congenital obstruction. The fertilization and pregnancy rates were not statistically different (p > 0.05) when testicular or epididymal spermatozoa were used in obstructive cases; neither was statistically different (p > 0.05) when compared in patients with congenital and acquired obstruction. On the other hand, the fertilization and pregnancy rates in cases with non-obstructive azoospermia were significantly lower (p < 0.05) than in obstructive cases.

Successful intracytoplasmic sperm injection without performing cytoplasmic aspiration.

OBJECTIVE: To compare the effect of cytoplasmic aspiration versus no aspiration before intracytoplasmic sperm injection (ICSI) on the rate of oocyte damage, fertilization rate, and embryo quality. DESIGN: A randomized prospective study on sibling oocytes. SETTING: The Egyptian IVF-ET Center, Cairo, Egypt. PARTICIPANTS: Fifty-eight patients who were infertile due to male factor who underwent 60 ICSI cycles. INTERVENTION: Intracytoplasmic sperm injection was performed on randomly allocated metaphase II oocytes with cytoplasmic aspiration in group I and without cytoplasmic aspiration in group II before sperm injection. MAIN OUTCOME MEASURE: Fertilization rate, oocyte damage rate, and embryo quality. RESULTS: Normal fertilization rate per injected oocyte was 61.4% in group I compared with 62.5% in group II. The damage rate per injected oocyte was 16.8% in group I compared with 4.6% in group II. Grade I embryos were 24.5% in group I compared with 48.5% in group II. CONCLUSION: Cytoplasmic aspiration before sperm injection in ICSI is not essential for oocyte activation. It did not improve the rate of normal fertilization. On the other hand, it increased the damaged oocyte rate and the rate of cytoplasmic fragments.

Schistosoma mansoni: identification and protective immunity of adult worm antigens recognized by T lymphocytes of outbred Swiss mice immunized with irradiated cercariae.

Percutaneous exposure of outbred Swiss mice to 500 Schistosoma mansoni (Egyptian strain) cercariae attenuated by 25,000 rad gamma radiation, twice at a 4-week interval, led to 80% protection against challenge with 100 live unattenuated cercariae compared to unimmunized control mice. The S. mansoni molecules that induce protective immunity in this model are not as yet identified. The capacity of an immunogen to induce efficient protective immunity depends largely on its T-cell-activating potential, as T cells are required both for eliciting long-lasting antibody formation and for antibody-independent cell-mediated immunity. To define such T cell antigen in S. mansoni, soluble adult worm antigens (SAWA) were separated by SDS-PAGE and electrotransferred onto nitrocellulose paper. Thirteen bands, identified by their M(r) were tested in T cell Western assays for their ability to stimulate proliferation of lymph node cells from 23 mice immunized with irradiated cercariae for the second time 2-5 weeks earlier. Lymphocytes from all mice responded to only a few (maximum of 6) bands. The response rate for the 13 SAWA bands tested ranged from 0-43%. These findings suggest a heterogeneity in T cell responses of individual mice to each of the SAWA bands and have implications that should be considered in the selection of immunogens to be assayed for anti-Schistosomiasis mansoni protective capacity. A highly significant protection against S. mansoni challenge in outbred Swiss mice was obtained exclusively following vaccination with a cocktail of soluble adult worm T cell immunogens that are recognized by 30-40% of individuals.