Search

The recruitment of radiolabelled blood-borne lymphocytes into rat popliteal lymph nodes (PLN) was investigated following the injection of varying doses of chicken (CRBC) or sheep red blood cells (SRBC) into the hind foot pad. Within the dose range tested (10(5)-10(8) RBC) an increase in the dose of injected antigen resulted in elevated levels of lymphocyte recruitment into the draining popliteal lymph node. The kinetics of lymphocyte recruitment with the same dose of CRBC or SRBC was similar even though these red cells differ in size and are antigenically non-cross-reactive. While little is known of the mechanisms which control the rate of entry of blood-borne lymphocytes into antigen-stimulated lymph nodes, the extent of lymphocyte recruitment was shown to be directly related to the quantity of antigen injected.

Orthodox views for the origin of the high concentration of prolactin (PRL) present in amniotic fluid suggest it is derived from maternal or fetal serum. However, the data on which these conclusions are based can also be interpreted to indicate that this hormone may be a product of placental or periplacental tissues. Trophoblast or amnion do not appear to produce PRL, while PRL synthesis by decidua-chorion is suggested from experiments in the rhesus monkey and by in vitro incubation of human tissue. Production of PRL by an extrapituitary cell is not without precedent and would be a simple explanation for high amniotic fluid PRL concentrations. Moreover, decidual-chorionic PRL would be strategically placed to mediate local functions of this hormone such as osmoregulation and myometrial inhibition.

The entry of radiolabeled blood-borne T and B lymphocytes into resting popliteal lymph nodes and popliteal lymph nodes stimulated with semiallogeneic lymphocytes was investigated in rats. Thoracic duct lymphocytes separated into T- and B-lymphocyte populations on nylon-wool columns were radiolabeled with 51chromium and equal numbers of T or B lymphocytes were injected intravenously. While the ratio of T and B lymphocytes in the blood is approximately 3:1 it was found that the ratio of T to B lymphocytes migrating into lymph nodes was approximately 9 T to 1 B lymphocyte in both resting and antigenically stimulated lymph nodes. Since the ratio of T to B lymphocytes in thoracic duct lymph is similar to that of blood, there is a disparity between the number of T cells entering and leaving lymph nodes. These results suggest that some T lymphocytes may return to the blood directly and/or there is increased T lymphocyte death in lymph nodes.

Using flow cytofluorimetry, the histamine-binding properties and T cell subclass distribution of peripheral blood lymphocytes in normal and atopic subjects were investigated. In the case of active (or symptomatic) atopy, the levels of histamine-binding lymphocytes were significantly reduced. However, none of these parameters distinguished between normal individuals and patient groups when the atopics were asymptomatic except the OKT3 marker; in the atopic group, the proportion of OKT3-positive cells was lower. These results indicate that the decreased levels of histamine-binding lymphocytes in atopy are a secondary phenomenon, and that the ratio of T helper to T suppressor/cytotoxic cells is not significantly altered in atopic patients. The changes in histamine-binding lymphocytes appear to be unrelated to the decreased numbers of OKT3-positive cells, since the latter changes were evident in active and asymptomatic atopics.

Immune reactivity to human keratoacanthoma was investigated by microcytotoxicity tests and immunofluorescence. IgM and complement were consistently present in lesions; IgG and fibrin were infrequent. No evidence of in vivo bound immunoglobulin was found on the surface of keratoacanthoma cells by membrane immunofluorescence. Neither patients' sera nor peripheral blood leukocytes showed significant cytotoxicity against autochthonous tumour cells in microtitre assays. This study fails to support the view that regression of human keratoacanthoma is mediated by immunological mechanisms.

A tubular or sac-like structure with a well defined lumen which occasionally contained parasites was found in the outer muscle wall of the small and large intestine in 32 out of 40 male and 35 of 40 female marsupial mice of the two species, Antechinus swainsonii and Antechinus stuartii (Macleay). A similar structure was also found in the muscle layers of the stomach in 5 marsupial mice of both species. The cellular matrix of the structure appeared to be composed of randomly organized macrophage-like cells and in one case there was evidence of an inflammatory reaction to parasites within the lumen. The origin of this tube is either vestigial or the result of reactive changes.

Langerhans' cells (LC) are the only subpopulation of epidermal cells to express the class II major histocompatibility (MHC) glycoproteins (H-2 Ia in the mouse) under normal conditions. Since these antigens are important in the initiation of allograft rejection, the effect of LC depletion on mouse skin graft survival was studied. Treatment with the chemical carcinogen 9, 10-dimethyl-1,2-benzanthracene (DMBA) was observed to deplete LC from the epidermis on ultrastructural examination. DMBA-treated C57BL dorsal trunk or tail skin grafted onto BALB/c recipients had a prolonged survival compared to solvent-treated donor skin. This was observed 1 week after a single DMBA treatment; successive once-weekly treatments or three treatments within 1 week failed to enhance allograft survival further. Tail-skin grafts had consistently longer survival times in comparison to dorsal trunk-skin grafts, for both control and treated skin. Treatment of dorsal trunk or tail skin with DMBA probably enhanced skin graft survival on allogeneic recipients by causing a loss of LC, and therefore of the class II MHC antigens from the graft. To confirm this, congenic mouse strains were used: B10.A(2R) x B10.A differing only at H-2D, and B10.A(2R) x B10.A(4R) differing only at H-2 I-E. Treatment of B10.A(2R) tail skin with DMBA for 1 week did not affect its survival when grafted onto H-2D-disparate B10.A mice, whereas when grafted onto H-2I-E-disparate B10.A(4R) hosts the DMBA-treated grafts were not only accepted permanently, but induced specific unresponsiveness. This confirms that the DMBA-induced enhancement of graft survival was the result of a loss of H-2 Ia antigens, and therefore of LC from the treated graft.

A seroepidemiological study of naturally occurring antibodies to the human syncytial virus has been carried out by means of an indirect immunofluorescence test on 639 East Africans, consisting of 493 normal Ugandans, 66 Kenyan patients with nasopharyngeal carcinoma (NPC), and 80 Kenyan and Tanzanian patients with various other tumours or non-cancerous conditions. It was found that 3.4% of the normal individuals had antibodies to the virus and serial serum samples were available from 14 of these, permitting the study of antibody class in seroconversion and antibody persistence. As in an earlier survey, a significantly higher incidence of antibodies was found amongst NPC patients. Blocking and indirect immunofluorescence test with simian foamy viruses (SFV) showed some cross-reactivity between SFV 6 and the human syncytial virus, but not identity. The results are discussed in relation to the very real occurrence of natural infection by human syncytial virus in certain geographical regions.

The chemical carcinogen 7,12-dimethylbenz (a) anthracene (DMBA) is a potent carcinogen which, when applied to the skin of BALB/c mice weekly for 7-8 weeks, causes the induction of macroscopically visible skin tumours. We report that DMBA also depletes Langerhans cells (LC) from treated skin; the number of cutaneous LC is reduced by nearly 50% 3 days after the first application of DMBA, and continues to decrease upon further treatment. After 7-8 weeks of DMBA application, while tumours are becoming macroscopically visible, there is a considerably lower LC density in treated skin. Upon cessation of the DMBA treatment, the LC repopulate the skin, returning to control values within 55-64 days. During this repopulation of the skin by LC, the tumours begin to decrease in size. Since LC function as local cutaneous antigen-presenting cells, and are responsible for initiation of an immune response against antigens in the skin, their depletion during tumour induction may allow DMBA-transformed cells to circumvent the immune system and form tumours. Their reappearance associated with tumour regression suggests that the LC are involved in an immune response against the tumours.

The exposure of murine skin to potent chemical carcinogens induced distinctive effects on the distribution of epidermal Langerhans cells (LC). Our previous finding that weekly applications of 7,12-dimethylbenz[a]anthracene deplete the numbers of adenosine triphosphatase (ATPase)-positive LC was extended to show that LC are also depleted on Ia and beta-glucuronidase staining. In contrast, application of the tobacco-derived carcinogen, benzo[a]pyrene (BP), caused a significant increase in Ia-positive LC density within 2 weeks and elevated levels were maintained for up to 6 months with continuous treatment. The tobacco-derived cocarcinogenic agent, catechol, also enhanced the numbers of epidermal LC. The LC in carcinogen treated epidermis were morphologically abnormal; after BP and catechol treatment LC appeared smaller with shorter dendrites, whereas in DMBA treated epidermis LC were enlarged with elongated dendrites. Application of the contact sensitizing agent, dinitrofluorobenzene, to skin treated with BP induced hyporesponsiveness rather than contact sensitivity upon subsequent antigen challenge. Hence, the function of the large number of morphologically altered LC in BP treated skin was impaired. We conclude that carcinogen-induced alterations of LC are associated with impaired immunocompetence, although different carcinogens probably operate via different mechanisms to induce such phenomena.

The basal lamina in a variety of skin tumours was assessed with an antibody to type IV collagen and the peroxidase-antiperoxidase (PAP) technique. The antibody was raised in rabbits against type IV collagen isolated from human placental tissue. The basal lamina in Bowen's disease was essentially intact while squamous cell and basal cell carcinomas showed focal loss in areas of tumour cell invasion. However, both tumours showed preservation of basal lamina around the majority of projections and nests of tumour within the dermis. Many keratoacanthomas showed extensive loss of the basal lamina. This loss appears associated with advanced keratinization at the base of the lesion and may represent an involutional change possibly secondary to inflammation. It is concluded that epidermal tumour cells following local invasion may cease migration and produce a distinct continuous basal lamina similar to that of the normal dermo-epidermal junction. Loss of basal lamina appears restricted to foci of ongoing tumour invasion.

Four human and 4 rat astrocytomata and mammalian adult brain and spinal cord were examined by indirect immunofluorescence with human serum containing smooth muscle antibody. Cryostat sections of astrocytomata showed staining of the tumour cell cytoplasm and processes while in normal adult brain and spinal cord the entire astrocyte stained. Impression film and tissue culture monolayers of astrocytomata showed staining of cell processes and a fine, filamentous network in the cell body. The reaction with astrocytoma tumour cells was stronger than that with the corresponding normal astrocytes. Specificity of the staining reaction was established by its prevention on neutralization absorptions of the serum with extracts or homogeneous of smooth muscle. The presence of smooth muscle-associated antigen in astrocytes and astrocytomata is indicative of contractile protein providing a mechanism of cell movement in vivo.

Cryostat sections of 29 squamous cell carcinomas, 13 keratoacanthomas, and 12 papillomas, induced by 7,12-dimethylbenz(a)anthracene in the skin of rabbits and rats were examined by indirect immunofluorescence with human serum containing antibody to smooth muscle. Linear or granular staining of the cell outlines of the basal squamous cell layers was seen most extensively in the carcinomas, less in keratoacanthomas, and least in papillomas. In addition, squamous cell carcinomas showed this pattern of staining at advancing tumor margins and in invasive cords and tumor cell nests in the dermis. Four keratoacanthomas also showed prominent staining of the basement membrane area. The specificity of the staining reaction was established by its prevention on neutralization absorptions of the serum with extracts or homogenates of smooth muscle. The epidermal cells of normal rabbit and rat skin gave negative staining reactions. The presence of smooth muscle-associated antigen probably corresponds to cellular microfilaments.

To study the nature of reticular epithelial (REp) cells and their role in the specific microenvironments of the chicken bursa and thymus, a method was developed for the in vitro culture of purified preparations of these cells. For comparison, similar cultures of splenic adherent cells were also performed. REp cell-rich bursa medullary follicles and mildly trypsinized thymic fragments were X-irradiated (850 rad) to eliminate remaining lymphocytes and transferred to culture flasks. In bursal cultures, after 2-4 days incubation the basement membrane (BM) encapsulating the follicles disrupted and the immediately underlying epithelial cells grew out as a monolayer. By 10 days, REp cells at the periphery developed cytoplasmic processes; occasionally these cells appeared to "bud-off" and grow as isolated dendritic cells. Thymic REp cells were generally slower to proliferate but formed a monolayer by 10-14 days. Splenic adherent cells developed extensive growth within 4 days. REp cells were distinguished from fibroblasts, when present, morphologically and by their limited phagocytic ability. The former were also periodic acid-Sciffs reagent (PAS)-positive and produced reticulin granules. Bursal REp cells were also positive for a gut-associated mucin, but this may have been bound in vivo prior to culture. Neither T nor B lymphocyte-specific antigens were detectable on the cultured REp cells or splenic adherent cells, but they were all rich in cytoplasmic actin. A major feature of REp cells to emerge in this study was the obvious presence of subpopulations of these cells, which raises important questions as to their exact nature and lineage. The accompanying paper details the ability of the bursal and thymic REp cell cultures to induce B-or T-lymphocyte differentiation, respectively, in vitro.

Lymph nodes from resected specimens of human colorectal carcinoma were investigated for in vitro lymphocyte cytotoxicity against primary cultures of autologous tumor cells. Regional lymph node lymphocytes were cytotoxic in 32 of 142 cases (23%). Altogether 200 nodes were examined and the cytotoxicity correlated directly with sinus histiocytosis, seen in 43 nodes from 35 cases, and with hyperplasia of B- and T-lymphocyte areas combined, seen in 92 nodes from 65 cases. Lymph nodes with combined B- and T-cell hyperplasia were significantly more common in cases of good tumor differentiation. The findings suggest that sinus histiocytosis and hyperplasia of both major lymphocyte populations are morphological expressions of in vitro antitumor immunoreactivity in the regional lymph node.

Cryostat sections of 16 preneoplastic and 14 neoplastic hepatic lesions induced in rats by 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB) were examined by indirect immunofluorescence with human serum containing smooth-muscle antibody (SMA). Preneoplastic lesions showed strong cytoplasmic staining of proliferating oval cells and of the cell outlines of hepatocytes, in areas of nodular hyperplasia. In carcinomas, poorly differentiated hepatocytes showed staining of cell outlines, while well differentiated tumour cells forming glandular structures showed only staining of the luminar surfaces. The stromal cells also showed cytoplasmic staining. Morphologically normal areas of 3'-Me-DAB-treated livers showed weak staining of cell outlines, similar to normal liver. Specificity of the staining reactions was established by failure of staining in parallel control sections treated with normal human serum, or SMA serum neutralized by absorptions with homogenates of smooth muscle or extracts of actin. The results suggest that there is an increased expression of actin-like contractile protein in preneoplastic and poorly differentiated neoplastic liver cells.

Langerhans' cell density was assessed quantitatively in cervical wart virus infection (cervical condyloma), cervical intraepithelial neoplasia, and koilocytic dysplasia with use of an antibody to S100 protein and an indirect immunoperoxidase technique. When compared with normal ectocervix, Langerhans' cell density was significantly decreased in cervical wart virus infection and significantly increased in cervical intraepithelial neoplasia. In koilocytic dysplasia, intermediate Langerhans' cell densities were obtained. In addition to being increased within the lesions of cervical intraepithelial neoplasia, Langerhans' cell density was increased in the adjacent normal ectocervix. Human papillomavirus, by reducing intraepithelial Langerhans' cell density, may decrease local immune surveillance and thus have a promoter effect in the development of cervical cancer. Following the development of cervical intraepithelial neoplasia the increase in intraepithelial Langerhans' cell density suggests a specific immune response directed against neoantigens associated with malignant transformation. If a permissive wart virus infection persists after transformation to cervical intraepithelial neoplasia (koilocytic dysplasia), continued depletion of Langerhans' cells results in intermediate densities.

Cryostat sections of livers from rats treated with carbon tetrachloride (CCl4) for 12 weeks were examined by indirect immunofluorescence for reactivity with human serum containing smooth muscle antibody (SMA). Increased binding of SMA serum to hepatocytes was observed as early as the first week of CCl4 treatment and remained enhanced during the 12 weeks of study. No difference in staining intensity was observed between pre-cirrhotic liver parenchymal cells and hepatocytes in regenerative nodules of frankly cirrhotic livers. In addition, granulation tissue fibroblasts in cirrhotic livers showed strong reactivity with SMA serum. The specificity of the staining reaction for actin was established by its prevention on neutralization absorptions of the serum with homogenates of smooth muscle or extracts containing actin derived from smooth muscle. Circulating SMA was not demonstrated in the serum of rats at any phase of the study. The observations show that there is an increased expression of actin-like protein in regenerating hepatocytes and granulation tissue fibroblasts in cirrhotic livers.

Eight out of nine patients with squamous cell carcinoma of skin have shown immunological reactivity against their own tumour cells by one or more tests with their sera or peripheral blood lymphocytes. The tests included membrane and cytoplasmic immunofluorescence, and, with cultured tumour, complement-dependent serum cytotoxicity and lymphocyte attack. One case examined in depth had an unusually conspicuous lymphocyte and plasma cell reaction on histological examination, and was positive by all four tests; a time-lapse cinephoto-micrographic record over seven days was obtained of the attack on the carcinoma cells in culture by the patient's lymphocytes.