Effects of pharmacological inhibition of hyaluronic acid synthesis on experimental endometriosis.

BACKGROUND: Dysregulated hyaluronic acid (HA) metabolism has been shown to be implicated in several pathologies including endometriosis. 4-Methylumbelliferone (4MU) is an HA synthesis inhibitor with proven antitumour activity. In this study, we aim to evaluate the effect of 4MU on endometriosis development both in vivo and in vitro. METHODS: Endometriosis was surgically induced by uterine tissue auto-transplantation in 32 two-month-old BALB/c mice. Animals were designated into the early or late starting treatment group, which initiated on day 2 or day 15 after surgery, respectively. Within each group, 4MU 200 mg/kg/day or vehicle (Control) were administered by oesophageal gavage for 28 days. After sacrifice, the percentage of developed lesions, lesion size, cell proliferation, vascularization and HA deposition within the endometriotic-like lesions were evaluated. Cell viability was assessed in endometrial epithelial cells (ECC-1) and in endometrial stromal cells (t-HESC); and migration was evaluated in t-HESC. RESULTS: There was a significant reduction in the percentage of developed lesions in mice that started the 4MU treatment on day 2 compared with its respective control group, and compared with those that started treatment on day 15. However, no significant changes were found when analysing endometriotic-like lesion's cell proliferation, vascularization and HA deposition. In vitro, both cell viability and migration were inhibited by 4MU treatment. CONCLUSIONS: The inhibition of HA synthesis could be a beneficial and alternative option to treat endometriosis at the early stage of the disease. Further research is necessary to elucidate 4MU's mechanism of action and better strategies for delivering this promising drug.

Resveratrol impairs cellular mechanisms associated with the pathogenesis of endometriosis.

RESEARCH QUESTION: Does resveratrol exert a potent inhibitory effect on the development of endometriosis by interfering with some pivotal processes? DESIGN: In-vitro cultures of primary endometriotic stromal cells, immortalized endometrial stromal (St-T1b) and endometriotic epithelial (12Z) cells were used to assess the effects of resveratrol on endometrial cell mechanisms. The effects of resveratrol on 12Z and St-T1b cell viability were assessed by MTT assay, apoptosis by FITC Annexin V assay and cleaved caspase-3 levels and cell migration by wound healing assay. The effect of resveratrol on the expression of genes related to cell migration, angiogenesis and cell stemness was evaluated by qRT-PCR. RESULTS: Resveratrol significantly decreased cell viability (P= 0.0065 to P = 0.0180), cell migration (P < 0.001 to P = 0.0225) and increased the number of apoptotic cells (P = 0.0031 to P = 0.0432) in both cell lines. In cell lines and primary culture, the treatment reduced MMP-2/TIMP-1 (P < 0.001 to P = 0.0180), VEGF (P = 0.0052 to P = 0.0243) and Ang-1 mRNA (P < 0.001 to P = 0.0382) expression. Among the stem cell phenotype markers, resveratrol 100 µM increased mRNA expression levels of Notch-1 (P < 0.001 to P = 0.0018), KLF-4 (P = 0.0011 to P = 0.0137), SOX-2 (P < 0.001 to P = 0.0070) and TERT (P < 0.001 to P = 0.0193) in both cell lines and primary cultures. The mRNA expression level of Snail-1 increased in the cell lines (P < 0.001 to P = 0.0087), whereas OCT-4 mRNA expression increased in St-T1b (P = 0.0396) and primary cultures (P = 0.0148). Vimentin mRNA expression showed a significant upregulation in primary cultures (P < 0.001). The expression of Msi-1 (P = 0.0145) and NANOG (P = 0.0080) decreased only in St-T1b cells. CONCLUSION: Resveratrol showed inhibitory effects on cell behaviour related to the development of endometriosis by differentially affecting growth, apoptosis, migration and stem cell phenotype of endometrial and endometriotic cells in vitro.

Effect of urolithins A and B on ectopic endometrial growth in a murine model of endometriosis.

Endometriosis is an often painful disease in reproductive-aged women, in which endometrial-like tissue grows outside the uterine cavity. Since the limited current therapeutic alternatives fail in alleviating the symptoms and based on our previous research in in vitro models using the same compounds as the ones used in the present study, we aimed to evaluate the effects of urolithins A (UA) and B (UB) on the growth and survival of endometriotic-like lesions in a murine model of endometriosis. Female BALB/C mice were surgically induced with endometriosis and treated with 2.5 mg kg-1 day-1 intraperitoneal UA or UB. The mice were monitored daily and weighed and the estrous stage was determined. After 28 days of treatment, lesions were counted, measured, excised, and fixed. Both urolithins proved not to affect the estrous cycle or body weight of the mice. UA completely prevented endometriotic-like lesions, while UB diminished the implant volume (p < 0.05). Treatment also reduced epithelial and stromal cell proliferation within the implants (p < 0.001 and p < 0.01, respectively) and apoptosis was enhanced (p < 0.05 and p < 0.01, respectively). These results are promising and reveal that urolithins A and B, separately, have a beneficial effect on the overall endometriotic growth without affecting the body weight or estrous cycle.

Interplay between Endometriosis and Pregnancy in a Mouse Model.

OBJECTIVES: To evaluate the effect of endometriosis on fertility and the levels of the IL-2 and IFN-γ in the peritoneal fluid in a mouse model; to evaluate the effect of pregnancy on endometriotic lesion growth, apoptosis and cell proliferation. STUDY DESIGN: Two month old C57BL/6 female mice underwent either a surgical procedure to induce endometriosis or a sham surgery. Four weeks after surgery mice were mated and sacrificed at day 18 of pregnancy. Number of implantation sites, fetuses and fetal weight were recorded. Endometriotic lesions were counted, measured, excised and fixed. Apoptosis and cell proliferation were evaluated in lesions by TUNEL and immunohistochemistry for PCNA respectively. Levels of IL-2 and IFN-γ were assessed by ELISA in the peritoneal fluid. RESULTS: Pregnancy rate (i.e. pregnant mice/N) decreased in mice with endometriosis. However there were no significant differences in resorption rate, litter size and pup weight between groups. IFN-γ augmented in endometriosis mice independently of pregnancy outcome. Additionally IFN-γ increased in pregnant endometriosis mice compared to pregnant sham animals. While IFN-γ increased in non pregnant versus pregnant mice in the sham group, IL-2 was increased in non pregnant mice in the endometriosis group. The size of endometriotic lesions increased in pregnant mice while apoptosis increased in the stroma and cell proliferation decreased in the epithelium of these lesions. Additionally, leukocyte infiltration, necrosis and decidualization were increased in the same lesions. CONCLUSIONS: Pregnancy rate is reduced in this mouse model of endometriosis. Levels of IL-2 are increased in the peritoneal fluid of mice with endometriosis suggesting a role of this cytokine in infertility related to this disease. The size of endometriotic lesions is increased in pregnant mice; however pregnancy has a beneficial effect on lesions by decreasing cell proliferation and by increasing apoptosis, decidualization and necrosis.

Anastrozole and celecoxib for endometriosis treatment, good to keep them apart?

Endometriosis is a benign gynecological disease. Cyclooxygenase-2 (COX-2) and aromatase proteins have been shown to be overexpressed in eutopic endometrium from women suffering from this disease compared to disease-free women. Furthermore, inhibition of these molecules individually was demonstrated to have antiproliferative and proapoptotic effects both in vitro and in vivo in several models. In this study, the effect of combining celecoxib, a selective COX-2 inhibitor, and anastrozole, an aromatase inhibitor, on the implantation and growth of endometriotic like lesions in a murine model of endometriosis was evaluated. Endometriosis was surgically induced in female BALB/c mice. After 28 days of treatment with celecoxib, anastrozole, or their combination, animals were killed and lesions were counted, measured, excised, and fixed. Immunohistochemistry for proliferating cell nuclear antigen and CD34 was performed for assessment of cell proliferation and vascularization. TUNEL technique was performed for apoptosis evaluation. Celecoxib was the only treatment to significantly reduce the number of lesions established per mouse, their size and vascularized area. In addition, cell proliferation was significantly diminished and apoptosis was significantly enhanced by both individual treatments. When the therapies were combined, they reversed their effects. These results confirm that celecoxib and anastrozole separately decrease endometriotic growth, but when combined they might have antagonizing effects.

[Immunology of pregnancy]. / Inmunología del embarazo.

One of the phenomena that offers more questions in Immunology is: Why the embryo, behaving like a semialogenic graft, is not rejected by the mother? It is known that the mother produces an active immunological response against the fetus and, nevertheless, in normal conditions, the immunological rejection does not take place. In the present work of revision, some mechanisms are described by means of which the specific immunological tolerance of the mother is generated towards the embryo. All these mechanisms are interdependent and altogether they constitute a safety network to avoid the fetal rejection. Here the effects of female sex hormones on the immunological system, the change of the profile of cytokines, the generation of immunomodulating proteins and blocking antibodies, the effect of the expression of the HLA-G and the paper of some cells, like Regulatory T lymphocytes, dendritic and Natural Killer cells, are detailed. Also other routes by which the embryo defends itself of the maternal immunological attack are described, like the induction of apoptosis in the endometrium and leukocytes, the tryptophan and iron metabolisms, the inhibition of the complement system and the expression of annexins.

The inhibitory effect of celecoxib and rosiglitazone on experimental endometriosis.

OBJECTIVE: To evaluate the effects of celecoxib and rosiglitazone on the implantation and growth of endometriotic-like lesions in a murine model of endometriosis. DESIGN: Prospective experimental study. SETTING: Animal research and laboratory facility. ANIMAL(S): Two-month-old female BALB/c mice. INTERVENTION(S): Surgically induced endometriosis in female BALB/C mice; 28 days of treatment with celecoxib, rosiglitazone, or their combination; counting, measuring, excising, and fixing lesions. MAIN OUTCOME MEASURE(S): Immunohistochemical examination for proliferating cell nuclear antigen (PCNA), CD31, and CD34 to assess cell proliferation and vascularization, with the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) technique for apoptosis evaluation. RESULT(S): Celecoxib and the combined treatment (celecoxib and rosiglitazone) statistically significantly reduced the mean number of lesions established per mouse, and all treatments diminished the implant volume. In addition, cell proliferation within the implants was statistically significantly reduced, and apoptosis was statistically significantly enhanced by all treatments. Also, we found that all treatments diminished the vascularized area in the lesion. CONCLUSION(S): These results are promising and reveal that celecoxib and rosiglitazone, combined or separately, have a beneficial effect on overall endometriotic growth.

Inmunología del embarazo / Immunology of pregnancy

Uno de los fenómenos que abre más interrogantes en la Inmunología es ¿Por qué el embrión, comportándose como un injerto semialogénico, no es rechazado por la madre? Se sabe que la madre produce una activa respuesta inmunológica frente al feto y sin embargo, en condiciones normales, el rechazo inmunológico no se produce. En el presente trabajo de revisión, se describen algunos mecanismos por medio de los cuales se genera la tolerancia inmunológica específica de la madre hacia el embrión. Todos estos mecanismos son interdependientes, y en conjunto constituyen una trama para evitar el rechazo fetal. Aquí se detalla la acción de las hormonas sexuales femeninas sobre el sistema inmunológico, el cambio del perfil de citoquinas, la generación de proteínas inmunomoduladoras y de anticuerpos bloqueantes, el efecto de la expresión de los HLA-G y el papel de algunas células inmunocompetentes como los linfocitos T reguladores, las células dendríticas y las células asesinas naturales o Natural Killer. Asimismo se detallan otras vías por las cuales el embrión se defiende del ataque inmunológico materno como la inducción de la apoptosis en el endometrio y en los leucocitos, el metabolismo de hierro y triptofano, la inhibición del sistema del complemento y la expresión de anexinas.

One of the phenomena that offers more questions in Immunology is: Why the embryo, behaving like a semialogenic graft, is not rejected by the mother? It is known that the mother produces an active immunological response against the fetus and, nevertheless, in normal conditions, the immunological rejection does not take place. In the present work of revision, some mechanisms are described by means of which the specific immunological tolerance of the mother is generated towards the embryo. All these mechanisms are interdependent and altogether they constitute a safety network to avoid the fetal rejection. Here the effects of female sex hormones on the immunological system, the change of the profile of cytokines, the generation of immunomodulating proteins and blocking antibodies, the effect of the expression of the HLA-G and the paper of some cells, like Regulatory T lymphocytes, dendritic and Natural Killer cells, are detailed. Also other routes by which the embryo defends itself of the maternal immunological attack are described, like the induction of apoptosis in the endometrium and leukocytes, the tryptophan and iron metabolisms, the inhibition of the complement system and the expression of annexins.

Effect of vascular endothelial growth factor inhibition on endometrial implant development in a murine model of endometriosis.

The main factor involved in neovascularization of ectopic endometrial tissue in endometriosis is the vascular endothelial growth factor (VEGF), which is produced both by the endometrial implant and by peritoneal macrophages. On the other hand, bevacizumab is an antiangiogenic agent used in the treatment of different tumors, like colorectal, pulmonary, and recently mammary. We evaluated the effect of the inhibition of VEGF activity with bevacizumab (Avastin) on ectopic endometrial growth in a murine model of endometriosis. Two months old female BALB/c mice had surgery performed to induce endometriotic-like lesions. Treatment with bevacizumab started on post-surgery day 15 and continued during 2 weeks. Then, animals were sacrificed, peritoneal fluid was collected, and endometriotic-like lesions were counted, measured, and removed. Cell proliferation, vascular density, and apoptosis were assessed by immunohistochemistry for proliferating cell nuclear antigen (PCNA), immunohistochemistry for CD34, and Terminal Deoxynucleotidil Transferase-Mediated dUTP Nick End Labeling (TUNEL), respectively. Vascular endothelial growth factor levels were evaluated in the peritoneal fluid by enzyme-linked immunoassay (ELISA). Treatment with bevacizumab significantly inhibited endometriotic lesion development (P < .05). Consistently, bevacizumab significantly inhibited cell proliferation in lesions (P < .01), reduced vascular density (P < .001), as well as increased the apoptotic cell percentage (P < .001). In addition, bevacizumab reduced VEGF levels in peritoneal fluid of endometriosis-induced animals (P < .05). In conclusion, this study suggests a direct effect of bevacizumab on the reduction of endometrial implant growth and supports further research on VEGF inhibition as a novel therapeutic modality in endometriosis.

Effect of vascular endothelial growth factor and interleukin-1beta on apoptosis in endometrial cell cultures from patients with endometriosis and controls.

The aim of this study was to evaluate the effect of vascular endothelial growth factor (VEGF) and interleukin-1beta (IL-1beta) on apoptosis induced by leuprolide acetate (LA) in endometrial epithelial cell cultures from patients with endometriosis. Primary endometrial epithelial cell cultures were obtained from uterine endometrial biopsies of patients with endometriosis and control women. Endometrial epithelial cells were incubated with LA; a combination of LA and VEGF; a combination of LA and IL-1beta; or in basal conditions. LA was added 3h prior to addition of VEGF and IL-1beta. After stimulation, the percentage of apoptotic cells was evaluated by the acridine orange-ethidium bromide technique and Bax expression was assessed by western blot. Treatment with LA enhanced the percentage of apoptotic cells in endometrial epithelial cells from subjects with endometriosis and control subjects. Addition of either VEGF or IL-1beta after exposure to LA restored the percentage of apoptotic cells to basal levels. Moreover, treatment with LA increased Bax expression in endometrial epithelial cells from patients with endometriosis. This effect was reverted by the addition of either VEGF or IL-1beta. Our results show that VEGF and IL-1beta reduce apoptosis and decrease Bax expression in endometrial epithelial cells from patients with endometriosis. This study suggests that VEGF and IL-1beta may protect endometriotic cells from undergoing apoptosis in addition to exerting their pro-angiogenic role.

Effect of aromatase inhibitors on ectopic endometrial growth and peritoneal environment in a mouse model of endometriosis.

OBJECTIVE: To evaluate the effect of aromatase inhibitors on ectopic endometrial growth and on the release of proangiogenic and proinflammatory factors in peritoneal fluid (PF). DESIGN: Prospective experimental study. SETTING: Animal research and laboratory facility. ANIMAL(S): Female Balb/c mice 2 months of age. INTERVENTION(S): Mice had surgery performed to induce endometriosis-like lesions. Treatment with anastrozole or letrozole was started on either postoperative day 1 or 28 and continued for 4 weeks. MAIN OUTCOME MEASURE(S): Endometriotic lesions were counted and measured and aromatase expression, cell proliferation, and apoptosis were assessed. Vascular endothelial growth factor (VEGF) and prostaglandin E (PGE) levels were evaluated in the PF. RESULT(S): Endometriosis-like lesions express aromatase P-450. Treatment with either anastrozole or letrozole did not prevent lesion establishment; however, it significantly decreased the size of the endometriotic lesion. When treatment was initiated on postoperative day 1, letrozole and anastrozole decreased cell proliferation and increased apoptosis. When treatment was started on postoperative day 28, both aromatase inhibitors decreased cell proliferation, but only anastrozole augmented apoptosis levels. In addition, letrozole reduced VEGF and PGE levels in PF. Anastrozole diminished VEGF content but did not cause any significant change in PGE levels. CONCLUSION(S): These findings support the further investigation of aromatase inhibition as a treatment option for endometriosis.

Hormonas sexuales y respuesta inmunológica / Sex hormones and immune system

Existe una evidente interrelación entre el sistema endócrino y el sistema inmunológico. Un ejemplo de esto es el efecto que las hormonas sexuales ejercen sobre las distintas poblaciones de leucocitos (linfocitos T y B, Células NK, granulocitos y macrófagos), así como sobre la producción y liberación de citoquinas y proteínas inmunoreguladoras. Tanto en las mujeres como en las hembras de otras especies, los estrógenos y la progesterona harían que primase una respuesta inmune humoral, lo cual resultaría beneficioso para la gestación, pero al mismo tiempo favorecería la aparición de ciertas enfermedades autoinmunes. Contrariamente, la testoterona haría que en los machos predominase la respuesta inmune celular. El siguiente trabajo es una revisión de distintos estudios referentes a la acción que las hormonas sexuales esteroideas ejercen sobre distintos componentes del sistema inmunológico.

Hormonas sexuales y respuesta inmunológica / Sex hormones and immune system

Existe una evidente interrelación entre el sistema endócrino y el sistema inmunológico. Un ejemplo de esto es el efecto que las hormonas sexuales ejercen sobre las distintas poblaciones de leucocitos (linfocitos T y B, Células NK, granulocitos y macrófagos), así como sobre la producción y liberación de citoquinas y proteínas inmunoreguladoras. Tanto en las mujeres como en las hembras de otras especies, los estrógenos y la progesterona harían que primase una respuesta inmune humoral, lo cual resultaría beneficioso para la gestación, pero al mismo tiempo favorecería la aparición de ciertas enfermedades autoinmunes. Contrariamente, la testoterona haría que en los machos predominase la respuesta inmune celular. El siguiente trabajo es una revisión de distintos estudios referentes a la acción que las hormonas sexuales esteroideas ejercen sobre distintos componentes del sistema inmunológico. (AU)

The role of GnRH analogues in endometriosis-associated apoptosis and angiogenesis.

It has been postulated that gonadotropin-releasing hormone (GnRH) analogues may act directly on endometrial cells and inhibit their growth and proliferation by regulation of apoptotic and angiogenic mechanisms. Eutopic endometrial cells from patients with endometriosis show an increased proliferation rate and are less susceptible to cell death by apoptosis than those from subjects without the disease. Notably, the GnRH analogue, leuprorelin, inhibits cell proliferation and increases the apoptotic rate in eutopic endometrial cell cultures, an effect that appears to be mediated by an increase in the expression of the pro-apoptotic proteins Bax and FasL and a decrease in the expression of the anti-apoptotic protein Bcl-2. Angiogenesis is an important process in the development of endometrial tissue, and it is regulated by vascular endothelial growth factors (VEGFs) and angiopoietins. VEGF levels are elevated in peritoneal fluid and endometriotic tissue from patients with endometriosis. In addition, it has been demonstrated that the expression of VEGF is potentiated by a variety of cytokines, including IL-1beta. Recent studies show that leuprorelin reduces the production of VEGF-A and IL-1beta in eutopic endometrial cell cultures, suggesting a mechanism by which it could inhibit the development of endometriosis. Thus, GnRH analogues appear to be effective in reducing the growth of endometrial cells, not only due to their classical pituitary endocrine effects, but also via a direct effect on the endometrial cells themselves.

Significance of ovarian macrophages in the follicular aspirates from ART patients.

PURPOSE: To evaluate the percentages of macrophages present in granulosa cells (GC) cultures from patients with different responses to the hyperstimulation, in relation to the percentages of apoptotic cells (ApC), as well as to the release of cytokines. METHODS: We studied 42 patients: 12 Hyporesponders, (with < or =4 follicles), 15 Normoresponders, (5-14 follicles), and 15 Hyperresponders, (> or =15 follicles). In GC cultures percentages of macrophages and ApC were counted and, in the conditioned media, cytokines were measured. RESULTS: Percentages of macrophages were significantly higher in GC cultures from Hyporesponders compared with Hyperresponders patients. Also, the percentages of ApC cells were the highest in Hyporesponders. On the contrary, cytokines concentrations were the lowest in this group. CONCLUSIONS: The low ovarian response is probably due to the decreased angiogenesis, which in turn produces increased apoptosis and decreased production of cytokines. The increased percentage of macrophages could be related to increased frequency of apoptotic cells.

Direct injection of vascular endothelial growth factor into the ovary of mice promotes follicular development.

OBJECTIVE: To investigate the effects of an ovarian injection of vascular endothelial growth factor (VEGF) on antral follicle development, neoangiogenesis, and apoptosis. DESIGN: Controlled laboratory study. SETTING: University-affiliated fertility center. ANIMAL(S): Balb/c female mice (n = 32) were studied. INTERVENTION(S): Mice were divided into four groups: control group (C) n = 6, no treatment; hyperstimulated group (HS), n = 8, ovaries were stimulated with 7.5 IU pregnant mare serum gonadotropin (PMSG) and 10 IU of hCG; VEGF group (V), n = 8, injected with 0.1 mL of VEGF (0.2 microg) in each ovary; V+HS, n = 8 injected with VEGF and 2 weeks later hyperstimulated. MAIN OUTCOME MEASURE(S): Number of antral and luteinized follicles, number of vessels, and percentage of Bcl-2-positive cells. RESULT(S): The number of antral follicles with VEGF was higher than in the C and HS groups (16.0 +/- 2.5 vs. 6.0 +/- 0.9 and 11.3 +/- 0.6, respectively, p<0.005). All treatments significantly increased the number of vessels (C: 5.0 +/- 0.5 vs. V: 20.0 +/- 4.8, p<0.005 and V+HS: 22.2 +/- 1.2, p<0.01), as well as increased Bcl-2-positive cells compared to controls (C: 0; V: 11.8 +/- 3.5, p<0.005; V+HS: 12.5 +/- 3.7, p<0.005). CONCLUSION(S): Our findings demonstrated that a direct injection of VEGF into the mouse ovary results in the development of an enhanced vascular network promoting follicular development and diminishing apoptosis.

Evaluacion de la producion de interleuquina 1 por celulas adherentes en pacientes con diabetes tipo II / Production of interleukin 1 by adherent cells in patients with Diabetes Mellitus type II

Se evaluó la actividad comitogénica de la interleuquina 1 producida en cultivos de células adherentes mononucleares de pacientes con diabetes mellitus (DM) tipo II o insulino no dependente. Dicha actividad fue medida por la incorporación de timidina en cultivos de timocitos de ratones (C3H/Hej, en presencia de fitohemaglutinina (PHA). Se observó que los sobrenadantes de los cultivos de células adherentes de los pacientes con DM tipo II no produjeron efecto sinergista con la PHA, obteniéndose niveles de linfoproliferación semejantes a los obtenidos en ausencia de dicha lectina. Esta falta de efecto comitogénico (p < 0,001) con respecto a la respuesta obtenida con la interleuquina 1 de sujetos normales más PHA, podría deberse a la liberación de algún/os factor/es solubles que podieron interferir con los receptores glicosilados para mitógenos o con los procesos de activación celular

Evaluacion de la producion de interleuquina 1 por celulas adherentes en pacientes con diabetes tipo II / Production of interleukin 1 by adherent cells in patients with Diabetes Mellitus type II

Se evaluó la actividad comitogénica de la interleuquina 1 producida en cultivos de células adherentes mononucleares de pacientes con diabetes mellitus (DM) tipo II o insulino no dependente. Dicha actividad fue medida por la incorporación de timidina en cultivos de timocitos de ratones (C3H/Hej, en presencia de fitohemaglutinina (PHA). Se observó que los sobrenadantes de los cultivos de células adherentes de los pacientes con DM tipo II no produjeron efecto sinergista con la PHA, obteniéndose niveles de linfoproliferación semejantes a los obtenidos en ausencia de dicha lectina. Esta falta de efecto comitogénico (p < 0,001) con respecto a la respuesta obtenida con la interleuquina 1 de sujetos normales más PHA, podría deberse a la liberación de algún/os factor/es solubles que podieron interferir con los receptores glicosilados para mitógenos o con los procesos de activación celular (AU)