Maintained allogeneic pregnancy in rats depleted of T cytotoxic/suppressor cells by OX8 monoclonal antibody treatment

It has been suggested that CD8 positive suppressor T-cells might be of importance in the non-rejection of the fetus. In the present investigation allogeneically pregnant (Lewix x DA) rats were subjected to in vivo treatment with monoclonal OX8 antibodies, reactive with the rat equivalent of CD8 receptor. This treatment protocol drastically reduced the numbers of OX8 positive cells in spleens and para-aortic lymph nodes. On the day of delivery these rats, together with normal IgG treated controls, were dissected and analysed for effects on: (1) fetal survival; (2) weight and immunohistology of spleens and para-aortic lymph nodes; (3) total numbers of IgM- and IgG-secreting cells within these organs. The OX8 treated rats passed through pregnancy as successfully as did the controls. Both groups showed the same type of pregnancy-induced changes in their lymphoid organs, including dramatic growth of the para-aortic lymph nodes and increase in Ig-secretors within both spleen and para-aortic lymph nodes. This pattern was the same in all pregnant rats investigated, including untreated syngeneically mated Lewis rats. It is concluded that OX8 positive T "suppressor" cells do not play an important role in the maintenance of the fetal-placental unit.     

An immunoscintigraphic evaluation of the engineered human monoclonal antibody (hCTMO1) for use in the treatment of ovarian carcinoma

Objective To assess the safety and targeting ability of the engineered human antibody (hCTMO1) in women with ovarian carcinoma.
Design The monoclonal antibody labelled with Indium-1 11 was administered to women with suspected primary or recurrent ovarian carcinoma six days pre-operatively. The first group of women was given a dose of 0.1 mg per kg body weight of radiolabelled antibody. A second group of women received 1 mg per kg body weight and finally a third group was given 1 mg per kg body weight of unlabelled antibody followed one hour later by 0.1 mg per kg body weight of radiolabelled antibody. All the women were then imaged using a gamma camera one hour and up to 96 hours after injection.
Participants Fourty-four women in whom there was a high suspicion of primary ovarian carcinoma on the basis of ultrasound or CT imaging and serum CA125 and those in whom there was a suspicion of recurrent ovarian carcinoma after being treated for histologically confirmed carcinoma.
Setting The Queen's Medical Centre, Nottingham and University Hospital Vrije Universiteit, Amsterdam, The Netherlands.
Results At the low dose of antibody the sensitivity for detection of ovarian carcinoma was 70%. After increasing the dose of antibody and also after pre-dosing with unlabelled antibody the sensitivity increased to 100%, but there was a large number of false positive results at the higher dose, and therefore the specificity was low. The liver and bone marrow were the organs with the highest activities.
Conclusion The genetically engineered antibody hCTMO1 is safe for use in women. This antibody effectively targets ovarian carcinoma and has greater potential as a vector for therapeutic use than as a diagnostic agent.     

An immunoscintigraphic evaluation of the engineered human monoclonal antibody (hCTMO1) for use in the treatment of ovarian carcinoma.

OBJECTIVE: To assess the safety and targeting ability of the engineered human antibody (hCTMO1) in women with ovarian carcinoma. DESIGN: The monoclonal antibody labelled with Indium-111 was administered to women with suspected primary or recurrent ovarian carcinoma six days pre-operatively. The first group of women was given a dose of 0.1 mg per kg body weight of radiolabelled antibody. A second group of women received 1 mg per kg body weight and finally a third group was given 1 mg per kg body weight of unlabelled antibody followed one hour later by 0.1 mg per kg body weight of radiolabelled antibody. All the women were then imaged using a gamma camera one hour and up to 96 hours after injection. PARTICIPANTS: Fourty-four women in whom there was a high suspicion of primary ovarian carcinoma on the basis of ultrasound or CT imaging and serum CA125 and those in whom there was a suspicion of recurrent ovarian carcinoma after being treated for histologically confirmed carcinoma. SETTING: The Queen's Medical Centre, Nottingham and University Hospital Vrije Universiteit, Amsterdam, The Netherlands. RESULTS: At the low dose of antibody the sensitivity for detection of ovarian carcinoma was 70%. After increasing the dose of antibody and also after pre-dosing with unlabelled antibody the sensitivity increased to 100%, but there was a large number of false positive results at the higher dose, and therefore the specificity was low. The liver and bone marrow were the organs with the highest activities. CONCLUSION: The genetically engineered antibody hCTMO1 is safe for use in women. This antibody effectively targets ovarian carcinoma and has greater potential as a vector for therapeutic use than as a diagnostic agent.     

Anticollagen antibody responses in DBA/1 (H-2q) mice associated with type II collagen-induced arthritis and the effect of pregnancy

We have previously shown that DBA/1 mice immunized with heterologous type II collagen showed remission of the subsequent collagen-induced arthritis (CIA) when pregnant, but experienced exacerbation postpartum. Measurement of anticollagen antibody (aCa) responses by ELISA in primiparous mice immunized at day 1 of pregnancy revealed no significant difference compared to aCa titres in virgin animals, apart from slightly increased titres following the primary immunisation. When mice received collagen challenge during early pregnancy, however, the date at which maximal antibody titres was reached was delayed by 5 days. Pregnancy initiated following the intraperitoneal boost caused a ten-fold suppression in aCa titres with a rise post-partum. Measurements of aCa levels in individuals which showed fetal resorption indicated that suppression of humoral responses was dependent on the presence of a viable conceptus. Antibody titres declined in all animals after a period of time, which was more prolonged in multigravidae where aCa titres were higher than in nulliparous and primiparous mice. The results show that although pregnancy alters aCa responses during the course of gestation, no long-term modification of humoral immunity occurs, an observation in agreement with the clinical findings in these mice and in humans.     

妊高征患者封闭抗体对配偶T淋巴细胞分化抗原CD3、CD4、CD8的影响

目的　通过测定妊娠高血压综合征 (妊高征 )患者血清封闭抗体对配偶T淋巴细胞的封闭效率来探讨妊高征发病的免疫学机制。　方法　正常孕妇组 82例 ,其中早孕组 16例 ,中孕组32例 ,晚孕组 34例 ;妊高征组 15例 (孕晚期 )。将孕妇血清与配偶T淋巴细胞作用后加入荧光标记的CD3、CD4、CD8单克隆抗体。以流式细胞仪测定所有研究对象的血清封闭抗体对配偶T淋巴细胞分化抗原CD3、CD4、CD8的封闭效率。　结果　正常孕妇组 ,早孕妇女对配偶T淋巴细胞CD3、CD4、CD8的封闭效率分别为 (4.14± 1.0 2、2 .0 2± 0 .2 4、2 .37± 1.0 5 ) % ,高于中孕期 (- 0 .2 9± 0 .13、1.0 3± 0 .2 7和 0 .6 5± 0 .2 3) % (P均 <0 .0 5 ) ;而晚孕期妇女血清对配偶T淋巴细胞CD3、CD4、CD8的封闭效率最低 (- 1.33± 1.4 7、0 .15± 0 .0 1和 - 1.0 4± 0 .37) %。妊高征患者血清对配偶CD3、CD4、CD8的封闭效率 (- 4 .16± 1.2 5、- 2 .13± 0 .4 3和 - 3.38± 1.0 6 ) %明显低于正常晚孕妇女 (P<0 .0 1、0 .0 0 5、0 .0 0 5 )。　结论　 (1)封闭抗体随孕周增加而降低 ,到晚孕期达到最低水平。 (2 )正常妊娠妇女孕早期血清高水平的封闭抗体 ,保证了胚胎早期的正常发育。(3)妊高征患者血清中封闭抗体的减少可能是导致妊高征发病的重     

Binding of alpha2 monoclonal antibody to human cervical tumor cell (SiHa) surface alpha2beta1 integrin modulates MMP-2 activity

OBJECTIVE(S): The purpose was to study the interrelationship between cell surface integrin receptor (alpha2beta1) and matrixmetalloproteinases. METHODS: Immunoprecipitation and cell adhesion assay were done to assay alpha2beta1 and alpha3beta1 on SiHa cell surface. Zymogram was developed to assay secreted MMP activity of cells grown in presence of alpha2 monoclonal antibody. Immunoblot was developed to assay expression of MMP-2, FAK, and p-FAK. Plasma membrane-dependent activation of MMP2 was performed by incubating pure MMP-2 with membrane-enriched fraction isolated from SiHa cells. RESULTS: Immunoprecipitation and cell adhesion assay results confirmed the presence of alpha2beta1 receptor on SiHa cells. Zymographic analysis of serum-free media collected at different time points from SiHa cells grown on alpha2 monoclonal antibody-coated culture dishes showed the expression and activation of MMP-2 within 2-4 h, confirmed by immunoblot. Western blot of cells grown on alpha2-coated dishes for 30 min-4 h showed increased phosphorylation of FAK. Membrane-enriched fraction isolated from SiHa cells was found to specifically activate proMMP-2 to its activated forms within 30 min. CONCLUSION(S): The experimental findings strongly indicate that SiHa cell surface alpha2beta1 regulates MMP-2 expression. Increased phosphorylation of focal adhesion kinase (FAK) strongly indicates the possible role of FAK in signaling cascade. Incubation of SiHa cell membrane fraction with pure MMP-2 strongly confirms the cell membrane-dependent activation of proMMP2.     

Langerhans cells in human papillomavirus (HPV) lesions of the uterine cervix identified by the monoclonal antibody OKT-6

The local inflammatory cell infiltrates in 263 cervical punch biopsies of the women followed-up since 1981 (16 +/- 14 months, mean +/- S.D.) for an established human papillomavirus (HPV) lesion with or without concomitant cervical intraepithelial neoplasia (CIN) were analysed for occurrence of Langerhans cells, defined by the monoclonal antibody OKT-6 using the avidin-biotin peroxidase complex (ABC) technique. OKT-6+ cells remained at the constant low level (1.5-1.9% of the inflammatory cells) in the different types of HPV lesions (flat, inverted or papillomatous condylomas), their percentages (range 0.8-2.1% of the cells) being slightly affected by the grade of HPV-associated CIN, however, (P less than 0.05 between HPV-CIN I and HPV-CIS). Although cervical HPV lesions characteristically are a disease of young females, the relative levels of in situ Langerhans cells did not show any age-dependence. Furthermore, the intensity of the inflammatory cell infiltrate did not correlate with the relative levels of OKT-6+ cells in the biopsies. Practically identical (1.6%) levels of OKT-6+ cells were found in the first biopsies of the HPV lesions shown to regress during the follow-up period (28.8% of cases), when compared with those (1.7%) in the lesions persisted (52.1% of cases) or progressed (19.1% of lesions). The results are discussed in terms of the proposed immune surveillance function against viral infections, attributed to Langerhans cells in the SALT/MALT concept.(ABSTRACT TRUNCATED AT 250 WORDS)     

Prevention of graft rejection by donor type II CD8(+) T cells (Tc2 cells) is not sufficient to improve engraftment in fetal transplantation

OBJECTIVES: Tc2 cells, a subset of CD8(+) T cells, are able to facilitate engraftment in a murine model of postnatal allogeneic bone marrow transplantation. The purpose of this study was to evaluate whether Tc2 cells could improve engraftment in fetal transplantation. METHODS: Gestational day 13 C57BL/6 (H-2(b)) fetal mice were used as recipients, adult B6D2F(1) mice (C57BL/6 x DBA/2, H-2(b/d)) as donors, and splenocytes from B6C3F(1) (C57BL/6 x C3H/He, H-2(b/k)) mice were used as stimulators in cultures used to generate the Tc2 cells from B6D2F(1) mice. Peripheral blood chimerism was examined monthly for 3 months. Thereafter, recipients were sacrificed to evaluate the levels of peritoneal, splenic and bone marrow chimerism. The T-cell responses of recipient splenocytes to cells of host origin were measured as a proliferative response in mixed lymphocyte cultures. RESULTS: Low levels of peripheral blood cell chimerism (<0.3%) were observed at 1 month of age, which declined further by 3 months of age. The levels of donor cells in the spleen, bone marrow and peritoneal cavity were usually not more than 0.05%. The peritoneal cavity tended to have higher levels of donor cells with 1 recipient sustaining as high as 25.03% at the age of 3 months. Higher peritoneal chimerism correlated with a lower donor-specific T-cell response. CONCLUSIONS: Transplantation of Tc2 cells was insufficient to improve bone marrow engraftment in utero, suggesting that graft rejection is not the major barrier to successful in utero transplantation. Donor cells can persist in the peritoneal cavity and might play an important role in inducing immune tolerance in fetuses.     

Anti-tumor activity of TRA-8 anti-death receptor 5 (DR5) monoclonal antibody in combination with chemotherapy and radiation therapy in a cervical cancer model

OBJECTIVES: There is substantial evidence that tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) causes apoptosis via activation of death receptors 4 and 5 (DR4 and DR5). We sought to determine the therapeutic potential of TRA-8 (anti-DR5 monoclonal antibody) in combination with chemotherapy and radiation therapy in a cervical cancer model. METHODS: DR5 expression in 7 human cervical cancer cell lines was analyzed by indirect immunofluorescence using murine TRA-8 in combination with flow cytometry. Cell lines were treated with TRA-8 alone or in combination with cisplatin, topotecan, or radiation, and cytotoxicity assays were performed. Mice were inoculated with ME-180 cancer cells and treated with different combinations of therapy. Animals receiving antibody were injected intraperitoneally with 200 microg of TRA-8. Animals received 9 Gy 60Co radiation divided into 3 fractions and 3 intraperitoneal doses of cisplatin (6 mg/kg) 1 h before radiation. A similar experiment was performed using topotecan (2 mg/kg) as the chemotherapeutic agent. RESULTS: DR5 was expressed to a varying degree on the cervical cancer cell lines. Combination treatment with TRA-8 and chemotherapy or radiation resulted in synergistic cytotoxicity in vitro. In vivo, combination therapy with TRA-8, cisplatin, and radiation produced tumor growth inhibition that was significantly greater than the other groups. Similar results were seen in combination studies with topotecan. CONCLUSIONS: These data suggest that DR5 is a good target for activation of the apoptotic pathway. Monoclonal antibodies such as TRA-8 may play an important role in the development of an effective treatment strategy for patients with advanced cervical cancer.     

Pharmacological treatment of severe hypertension in pregnancy and the role of serotonin(2)-receptor blockers

Hypertensive disorders of pregnancy are the leading cause of maternal and perinatal mortality and morbidity in developing and developed countries. The etiology of preeclampsia is still unknown. Delivering the baby is the only definite treatment. The benefits of acute pharmacological control of severe hypertension prior to and/or post-delivery are generally accepted. Most drugs commonly used in the management of severe hypertension in pregnancy have significant maternal and/or neonatal adverse side effects. Furthermore, some are not effective to acutely lower the blood pressure in patients with a hypertensive crisis. Until recently not one of the commonly used antihypertensive drugs has been tailored to the pathophysiology of severe preeclampsia, being a clinical syndrome characterized by endothelial cell dysfunction, vasospasm and platelet aggregation. Ketanserin, a serotonin(2)-receptor blocker, is a drug that appears to be tailored for treating this pregnancy-associated enthothelial cell dysfunction. The results of several prospective trials show that there is a definite place for serotonin(2)-receptor blockers in the treatment of pregnancy-induced hypertensive disorders. This review provides a summary on the more established drugs as well as on some of the newer antihypertensive drugs used in pregnancy with emphasis on the existing experience with ketanserin.     

Identification of human chorionic gonadotropin (HCG) secreting cells and other cell types using antibody to HCG and a new monoclonal antibody (mABlu-5) in cultures of human placental villi

Summary We have identified cells which secrete human chorionic gonadotropin (HCG) of cultures if first trimester placental villi. As a first step, we identified epithelial cells using a new monoclonal antibody. We then added HCG antibodies to the cultured cells. We found that syncytiotrophoblast (and not cytotrophoblast), Hofbauer cells and some mesenchymal cells stained with HCG antibodies.     

Localization of antigen defined by a monoclonal antibody to human 32-kDa insulin-like growth factor-binding protein in the sheep uterus at preimplantation stages of pregnancy.

Employing a murine monoclonal antibody to pregnancy-associated endometrial alpha I-globulin, a 32-kDa insulin-like, growth factor-binding protein (IGF-BP) and major product of the decidualized endometrium in the human, reactivity has been detected in the luminal epithelium of the sheep endometrium during the preimplantation period of pregnancy. Staining was associated with supranuclear regions of the cytoplasm. Reactivity was restricted to the pregnant uterus and was distributed throughout the luminal epithelium in both caruncular and intercaruncular regions. The reactivity was absent from endometrial glands in intercaruncular regions. Staining was first detected at day 10, peaked at day 14 of gestation and was absent by day 16. The implications of the induction of the production of this protein by the preimplantation blastocyst in the luminal epithelium is discussed with reference to the potential role of IGF-BP in IGF action in the embryo.     

Isolation and characterisation of a subpopulation of human chorionic cytotrophoblast using a monoclonal anti-trophoblast antibody (NDOG2) in flow cytometry.

Human cytotrophoblast cells, isolated from term amniochorion by enzymic digestion and Percoll gradient centrifugation, were characterised by flow cytometry. A panel of 12 anti-trophoblast monoclonal antibodies was screened for labelling of these cells in flow cytometry and the results compared with immunoperoxidase labelling of cytospin preparations and tissue sections. All 12 antibodies were positive for trophoblast on tissue sections, 11/12 were positive on cytospins but only two (NDOG2 and GB25) gave consistent results in flow cytometry. Two-colour labelling with NDOG2 and W6/32, an antibody to HLA-A, -B, -C, demonstrated that 88% of the NDOG2-positive cells also express Class I major histocompatibility complex (MHC) antigens. The NDOG2-positive cytotrophoblast subpopulation was isolated by flow cytometry in sufficient purity (greater than 95%) and yield (3.1 x 10(6)) for use in functional studies in vitro.     

The effect on pregnancy of treatment with antibodies against pregnancy-associated murine protein I (PAMP-I) in inbred and outbred mice

Intravenous injection of small amounts of monospecific rabbit IgG against pregnancy-associated murine protein I (PAMP-I) induced abortion in mice where there was a histocompatibility difference between mother and fetuses. No abortion could be induced in inbred mice by a similar treatment. The maternal serum level was found to be higher in inbred than in outbred mice. The abortive dose of antibodies did not influence the serum levels of PAMP-I. Histological examination of uterine, placental and liver tissue showed only morphological changes in the placental tissue of mice which aborted due to the treatment with anti-PAMP-I antibodies.     

Levels of serum amyloid P-component associated with pregnancy and collagen-induced arthritis in DBA/1 (H-2q) mice

The levels of the acute-phase reactant serum amyloid P-component (SAP) were measured by quantitative rocket immunoelectrophoresis in pregnant and non-pregnant DBA/1 female mice with or without collagen-induced rheumatoid arthritis (CIA). Non-pregnant animals with CIA showed elevated SAP titres related to the severity of the disease. Pregnancy alone also caused increased SAP levels equivalent to those found in animals with established CIA but which were virgin. The clinical remission seen in arthritic animals during pregnancy was not associated with reductions in circulating SAP levels. Increasing parity, however, caused a lowering of SAP levels in animals with or without CIA compared to the primiparous individuals. Pregnancy causes a strain-dependent elevation of serum SAP which is not further elevated by CIA, thus limiting the usefulness of SAP measurements in assessment of disease progression or remission during gestation.     

Immunological responses in pregnancy. The studies on the role of the uterus in gestation and effects to lymphocytes subpopulation (author's transl)

The lymphocytes that isolated from the both peripheral venous blood (PVB) and uterine venous blood (UVB) of pregnant women with a complication of myoma were studied on the subpopulation (E-rosette and S-Ig method) and lymphoblastoid transformation to mitogens. Two methods on the E-rosette in this study were applied to make a clear correlation between masking phenomenon and humoral factors. One is to be used lymphocytes that washed five times with P.B.S., the other one cultured in RPMI 1640 with 20% FCS for 72 hours afterward. The results were obtained as follows. 1) Although T-cell of UVB displayed significantly lower than that of PVB. The count of T-cell of both PVB and UVB elevated to standard level after washing five times with P.B.S. and/or cultured for 72 hours. 2) The average counts of B-cell by the S-Ig were not detected on the difference between of PVB and UVB. 3) No differences in ratio of lymphoblastoid were recognized on the both lymphocytes of PVB and UVB to PHA and PWM too. 4) AFter five times washing, the counts of T-cell in UVB improved to standard value range. Therefore no difference of the counts in T-cell were detected between in PVB and UVB.