W. Page Faulk

PREVENTION OF RECURRENT ABORTION WITH LEUCOCYTE TRANSFUSIONS

Abstract Three women, each with a history of three spontaneous abortions, were typed for A, B, C, and DR histocompatibility (HLA) antigens and found to share antigens with their husbands. The women were repeatedly transfused throughout pregnancy with leucocyte-enriched plasma from at least sixteen different erythrocyte-compatible donors. The pregnancies were normal and each mother produced a healthy baby. The presence of trophoblast/lymphocyte cross-reactive (TLX) antigens, which stimulate the mother to mount a response with blastocyst protective factors and which prevent maternal rejection of the antigenically unique embryo, might explain these results.

TRANSFERRIN AND TRANSFERRIN RECEPTORS IN CARCINOMA OF THE BREAST

Breast tissues from 59 patients were immunohistologically studied for transferrin and transferrin receptors. None of the 8 normal breasts or 22 benign mastopathies, and only 1 of 7 fibroadenomas were reactive for transferrin, but 16 (72.7%) of 22 carcinomas were positive and parallel sections showed histopathologically that the rective areas were confined to tumour-cell membranes. That breast-tumour transferrin is probably receptor-bound in vivo was indicatd by its complete chemical removal in vitro and return to the same cells following reincubation with dilute solutions of transferrin. Similar bindig is found on certain normal or transformed cells and at the maternofetal interface of human placentae. It is suggested that this oncofetal receptor preferentially binds apotransferrin and may offer a novel approach to more specific therapy of certain breast cancers.

HUMAN AMNION AS AN ADJUNCT IN WOUND HEALING

Biopsy specimens from the beds of leg ulcers of fifteen patients were obtained before and after the application for 5 days of cultured human amnion. After amnion application there was considerable granulation tissue in the ulcer bed and microscopical evidence of thinned connective tissues, vessel development, more compact resolution of vascular basement membranes, and many more factor VIII granules within endothelial cells. These findings suggest the presence of angiogenic factors in human amnion and this could explain the hitherto unexplained success of amniotic membranes in surgical practice.

Propidium iodide as a nuclear marker in immunofluorescence. II. Use with cellular identification and viability studies

The use of a fluorescence method employing propidium iodide to determine nuclear morphology of different cell types by epi-illumination in membrane immunofluorescence is described. Its usefulness is illustrated by differentiating nucleated and anuclear transferrin receptor-bearing cells as well as by simultaneously determining the viability of cell suspension without requiring a change from fluorescence microscopy, this causing no loss of visual accommodation.

The human materno-foetal relationship in malaria. II. Histological, ultrastructural and immunopathological studies of the placenta

Histological and ultrastructural studies of four placentae heavily infectd with Plasmodium falciparum revealed large intervillous accumulations of erythrocytes containing parasites together with monocytes which had ingested pigment. These appearances were associated with focal syncytial necrosis, loss of syncytial microvilli and proliferation of cytotrophoblastic cells. In addition, marked irregular thickening of trophoblastic basement membranes and protrusion of tongue-like projections of syncytiotrophoblast into the basement membrane were observed. In six other placentae which contained scanty amounts of pigment but no parasites, representing past or inactive infection, no large collections of monocytes or abnormalities of trophoblast were apparent but basement membrane thickening was evident. Immunohistological studies revealed no significant differences between placentae positive for parasites and those containing pigment only, although the amount of certain immunoproteins and clotting factors was clearly increased above normal. These findings establish that P. falciparum infection in the placenta may result in substantial damage although lesions within the villus are rare. Furthermore, previous infection, although adequately controlled, may leave a heritage of pigment deposition, basement membrane thickening and immunopathological lesions. These results may thus account for both the high frequency of intra-uterine growth retardation and the rarity of congenital malaria in the presence of P. falciparum malaria.

TREATMENT OF CHRONIC ULCERATION OF THE LEGS WITH HUMAN AMNION

Chronic leg ulcers in fifteen patients were prepared for autografting with a dressing of cultured human amnion. In thirteen vascular responses filled the ulcer beds with granulation tissue. Four patients initially received more traditional therapy and their lesions altered little in gross appearance and were not regarded clinically to have produced a granulation-tissue response. Fourteen patients received autografts after 5 days of amnion application and eight of these grafts have remained intact for 2-10 months. The appearance of healthy granulation tissue after amnion application was associated with successful autografting.

Allotypic trophoblast—lymphocyte cross-reactive (TLX) cell surface antigens

Evidence id presented for the presence of trophoblast-lymphocyte cross-reaction (TLX) cell surface antigens. Complement-dependent lymphocytotoxicity studies with antitrophoblast sera (Ce) raised to ten separate placentas showed variable reaction patterns on peripheral blood leukocytes isolated from 30 random HLA-typed donors. No correlation with any known antigens of the major histocompatibility complex was evident. Absorption studies with chromatographically prepared fractions of solubilized trophoblast membranes from individual placentas and absorptions with both normal and transformed lymphoid cells removed cytotoxicity from some but not other Ce-antisera when tested on a single donor, thus providing serological evidence for allotypic variations among the TLX antigens.

Loss of transferrin receptors following induced differentiation of HL-60 promyelocytic leukemia cells

Abstract Human promyelocytic leukemia (HL-60) and lymphoblastoid (Daudi) cells were studied: for transferrin receptors before and after induced differentiation with dimethyl sulfoxide (DMSO), sodium butyrate or retinoic acid. None of these reagents affected the morphology or presentation of receptors in Daudi cells, but many HL-60 morphologically matured to banded neutrophils and demonstrated a concomitant loss of transferrin binding, suggesting an important role for transferrin receptors in cellular differentiation.

MATERNAL BLOCKING FACTORS IN HUMAN PREGNANCY ARE FOUND IN PLASMA NOT SERUM

Pregnancy plasma and sera were evaluated for differential blocking effects in mixed lymphocyte culture reactions. Autologous pregnancy plasma, but not serum was found to contain a specific allogeneic inhibitor for paternal lymphocytes.

Cultivation of microvascular endothelial cells from human preputial skin

SummaryA procedure is described for the isolation and cultivation of microvascular endothelium from human skin. Neonatal foreskins are pooled, washed, minced, and dissociated by a mixture of collagenase and dispase. Microvascular endothelium, liberated in the form of intact capillary fragments, is incompletely separated from fibroblasts and epidermal cells by sieving through nylon mesh, followed by velocity sedimentation on 5% bovine serum albumin. The endothelium-enriched fraction has been maintained in primary culture for up to 3 weeks. The resulting epithelioid colonies have been characterized morphologically by both light and transmission electron microscopy and manifest all of the structural features that distinguish other, large-vessel endothelia in culture. In addition, immunohistochemical studies using an indirect fluorescent antibody technique demonstrate that these cells contain the endothelium-specific product, Factor VIII antigen.