Helena Tlaskalová

The phagocytic activity of reticuloendothelial system of newborn precolostral pigs to smooth and roughEscherichia coli

The fate of smooth and rough strains ofEscherichia coli in the blood stream of newborn, germ-free, colostrum deprived piglets was studied using blood clearance test with live and radioisotope-labelled (P32) bacteria.Smooth strains are eliminated at an extremly slow rate (hours) whereas rough strains are taken up within 30 minutes. Specific immune serum accelerates rapidly the clearance rate of smooth strains after bothin vitro andin vivo opsonisation of bacteria. Clearance of smooth liveEscherichia coli from the blood stream cannot be stimulated by an endotoxin treatment of newborn piglets. The possible role of complement system as a nonspecific opsonin in phagocytosis of rough strains is discussed. The results demonstrate that RES cells of newborn precolostral piglets are functionally competent as to the capacity to remove gramnegative bacteria from the blood stream provided specific and/or nonspecific opsonins are available.

Receptor-specific fractionation of immunocompetent cells and purification of antibodies on hydroxyalkyl methacrylate immunosorbents.

Abstract Immunosorbents prepared with hydroxyalkyl methacrylate gels activated by cyanogen bromide, have a binding capacity for specific antibodies from immune sera similar to that of commercial Sepharose immunosorbents. Moreover, they can be used in specific fractionation of immunocompetent cells. Enriched cells suspensions of antibody producing spleen cells from rabbits immunized with SRBC or HSA were obtained by using large gel particles (500–700 μm). Using the batch method, the antibody producing cells could be concentrated 5–30 times. Blocking of cell receptors with anti-immunoglobulin serum or specific antigen confirmed the specificity of cell attachment to the immunosorbent. According to the results obtained hydroxyalkyl methacrylate gels are suitable for concentration of memory cells and separation of T and B cells.

Isolation and properties of antigenic-specific receptors from rabbit and pig lymphoid cells

Abstract In order to study the character and properties of antigen-specific receptors on lymphocyte surfaces, an immunoadsorption method based on the principle described by Kiefer (1973, 1975) has been developed. This method permits isolation of receptors from rabbit and pig spleen and lymph node cells in quantities sufficient for basic analyses. Isolated receptors represent a mixture of two groups of antigen binding molecules. Both groups react with corresponding anti-idiotypic antisera but only one of them reacts with antisera to serum immunoglobulins. These two fractions can be separated using an immunoadsorbent with bound antibodies to serum immunoglobulins. In the fraction bearing immunoglobulin determinants, the presence of IgM and IgG but not IgA has been detected. The IgG receptors and IgG antibodies in the serum of the same animal had different isoelectrofocusing spectra. The origin of the other antigen-binding fraction remains to be established. The results obtained with antisera to the ‘constant part’ of this molecule confirmed that it does not carry conventional immunoglobulin determinants.

Ontogenetic development of antibody formation in response to different doses of gram-negative microorganisms in young rabbits. I. Dynamics and site of antibody formation after primary immunization.

Rabbits at different stages of development were immunized with different doses of heatinactivated suspension ofEscherichia coli 086 andSalmonella paratyphi B. The dynamics and the site of formation of bactericidal and haemolytic antibodies during the primary reaction was investigated. An increase and an acceleration of antibody formation after increasing the dose of antigen was found in the serum and at the cellular level. The magnitude of the response and the rate of the reaction were directly proportional to the age of the experimental rabbits. The site of antibody formation depends on the character, route of administration, antigen dose and age of rabbits. After intraperitoneal and also after intravenous immunization withEscherichia coli andSalmonella paratyphi B antigens the site of antibody production in 15-day-old rabbits was the lymphatic tissue of the intestine, the appendix, and mesenteric lymph nodes. As the antigen dose was increased and the age of rabbits rose, i.e. in correlation with the increase of the antibody response, antibody formation shifted to the spleen which is the chief site of antibody production following immunization by these bacterial antigens from the first month of life of rabbits. In contrast with this type of antigen, after intraperitoneal or intravenous immunization with sheep erythrocytes of new-born or older rabbits antibody formation was concentrated in the spleen. The development of the immunological competence and the significance of intestinal lymphatic tissue as one of peripheral type is discussed.

Some properties of piglet, calf, pig and bovine conglutinin.

In sera of newborn piglets which were prevented from sucking maternal colostrum a low titre of conglutinin activity was demonstrated. For comparison of properties of this piglet conglutinin sera derived from adult pigs, cows and calves were used. Conglutinin from precolostral piglet serum behaves in different way as compared with immunoconglutinin from adult pig following reduction with 2-mercaptoethanol and in reaction with EDTA and thus resembled bovine natural conglutinin. In density gradient ultracentrifugation and inhibition reaction with zymosan andn-acetyl-d-glucosamine reacts piglet conglutinin as pig immunoconglutinin. Conglutinin present in precolostral calf serum behaves like a typical natural conglutinin of adult cattle.

Detection of mouse cells producing antibodies against the azophenylarsonic group by haemolytic plaque assay

For detection of mouse cells producing anti-ARS antibody a modified method using ARS-SRBC is described. For optimal results a suitable source of SRBC must be selected. The number of ARS groups per SRBC required for optimal lysis in PFC assay varies, and ARS-SRBC prepared by the method of Ingraham which are suitable for detection of rabbit-PFC give negative results with the mouse system. ARS-SRBC prepared for the PFC assay are unsuitable for the haemagglutination test and vice versa.

Malignancy in Coeliac Disease and Dermatitis Herpetiformis

Publisher Summary Celiac disease (CD), gluten-sensitive enteropathy, is defined as a permanent intolerance of dietary gluten and similar proteins of wheat, rye, barley and oats. The disease is characterized by mucosal injury appearing in genetically susceptible individuals after gluten ingestion. A long-life gluten-free diet is used for treatment; it usually improves the morphological picture and clinical symptoms. The undiagnosed and/or untreated CD (patients not keeping a gluten-free diet) develops lymphoma or carcinoma. In untreated CD, there is a 43-fold higher relative risk of small bowel lymphoma, a 12-fold higher relative risk of cancer of the esophagus and a 10-fold higher relative risk of cancer of the mouth and pharynx. The chapter discusses various studies to a higher incidence of non-Hodgkins lymphoma in celiac patients than in general population. Although the pathogenic mechanism of CD is not yet fully elucidated, there is no doubt about the involvement of immune factors in the development of the disease. The chapter describes the various factors that link CD to autoimmune diseases. As with CD, an increased incidence of malignant disease (lymphoma) has been reported in patients with dermatitis herpetiformis. Dermatitis herpetiformis is a relatively rare skin disease characterized by a rash with small blisters and intense itch. The chapter briefly describes the causating factors, clinical symptoms, and pathogenesis of this disease. Cancer of the small intestine is rare as compared with other sites in the gastrointestinal tract. Out of the four major primary small bowel tumors (adenocarcinomas, lymphomas, carcinoid, and leiomyosarcomas), adenocarcinomas and lymphomas are associated with diseases that increase the risk of developing these malignancies. Therefore, treatment of the predisposing condition decreases the risk of developing subsequent malignancy.

Ontogenetic development of antibody formation in response to different doses of gram-negative microorganisms in young rabbits

Newborn and 15-day-old rabbits were immunized with different doses of heat-inactivated suspension ofEscherichia coli andSalmonella paralyphi B. The secondary immunization was performed after 4 weeks and the dynamics, magnitude and site of the secondary response studied. The magnitude of the secondary response was found to depend on the magnitude and rate of the primary response, this latter reflecting the dose used. A direct relationship was found in the range of the minimal and optimal dose: the higher the primary dose the higher the secondary response. After a rapid and pronounced primary response evoked by a high dose of 1010 microorganisms in 15-day-old rabbits, a partial inhibition of the secondary response was observed. The inhibition was pronounced primarily in the spleen; the lymphatic nodes reacted by a higher number of antibody producing cells as compared with the control nonprimed young rabbits.

Effects of Nocardia-delipidated cell mitogen on intestinal mucosa and spleen lymphocytes of germ-free rats.

Many compounds of bacterial origin can modulate basic physiological parameters of the mammalian organism including the immune system. One of them is Nocardia- delipidated cell mitogen (NDCM) which was isolated by delipidation from Nocardia opaca.1 NDCM stimulates proliferation of small resting human B lymphocytes and their differentiation into Ig-secreting cells.2 The mucosa of small intestine, especially the enterocytes, cells with digestive and absorptive function, produce a number of glycohydrolases. The disaccharidase (sucrase, lactase, and glucoamylase) activities of brush border membrane vesicles (BBMV) of enterocytes after a short-term NDCM- treatment have not been studied. Measurement of lymphocyte proliferation is an established method of quantifying the immune response to foreign antigens. Antigenic stimulation of human peripheral blood lymphocytes by NDCM was measured by Barot-Ciorbaru.2 Neither 3H -TdR-nor 3H-UdR-uptake by T cells has been measured after NDCM stimulation.

Stimulation of Galt by Nocardia Delipidated Cell Mitogen (NDCM) in Irradiated Germfree Piglets

Ionizing radiation seriously damages gastrointestinal compartments of the mucosal immune system. Even small doses lead to mitotic disturbances, death of stem cells in crypts of Lieberkuhn, injury of the epithelium and endothelium and extensive edema and hemorrhages. Low doses of ionizing radiation affect mostly lymphocytes which are extremely radiosensitive cells. Their morphology is influenced by doses as low as 0.25 Gy after 2 h. Lymphocytes which are in the interphase of cell cycle are killed after irradiaton by interphase death, a kind of programmed cell death.