AE rosette test (Wybran and Fudenberg) detects T lymphocyte subpopulation and correlates with cellular immunity. In this study we found decreased percentage of active T lymphocytes mainly in peripheral blood of newborns, in children with frequent infections and with malabsorption syndrome. We tried to correlate AE and E rosette tests with PHA lymphocyte reactivity in vitro in 231 patients. We found that decreased PHA lymphocytes reactivity better correlates with decreased percentage of active T lymphocytes than with total number of T lymphocytes. Active rosette test (AE) can be usefull in evaluation of cellular immunity in children.

The study of the ontogenesis of immunity in man is still in progress and development. In this paper the two major subpopulations of human lymphocytes--T (thymus dependent) and B (bursal equivalent or thymus independent) were examined during the ontogenetic development of man. These subpopulations can be examined in vitro by the methods distinguishing superficial receptors on these cells--namely by the rosseting methods. B lymphocytes with the receptor for Fc of immunoglobulin can be distinguished by the EA test, and with receptor for C3 by the the EAC test. T lymphocytes can be detected in blood by the E test using their capacity to bind sheep red cells forming rossetes. This phenomena is very dependent upon temperature changes during the experimental procedures and this may partly explain the widely different proportions of E rosseting forming cells found by different authors ranging from 37% to 81% in blood of healthy adults. So there is a great necessity for each laboratory to have their own normal value in the healthy children in different age to compare it with those estimated in different states of the disease. Percentages of T and B lymphocytes were evaluated in children in 5 different age groups: below 1 year, 2-3 years, 4-5 years and over 10 years. Moreover the number of T and B lymphocytes was evaluated in the peripheral blood of pregnant women (2 and 3 trimester), of women at delivery, in the cord blood and the blood of newborns on 5-th day of life. The normal control group consisted of 71 healthy adults. In some children of four age groups the absolute counts of T and lymphocytes were estimated. The results are presented in Tabl. I, II and. fig. 1, 2. We found significantly decreased percentage of T lymphocytes in some estimated groups versus control--it is in peripheral blood of pregnant women, women at delivery, in cord blood, in newborns at 5-th day of life (p less than 0.001) and in the groups of children below 5 years (0,01 less than p less than 0,02 and 0,02 less than p less than or equal to 0,05). In children older than 5 years the number of T lymphocytes was within normal limits. Total number of peripheral T lymphocytes was significantly increased in children age 2-5 years (p less than 0.001) and in children below 10 years (0.02 less than p less than or equal to 0.05). Percent of B lymphocytes estimated by the EA and EAC tests was elevated in pregnant women and women at delivery. The statistical difference between these two groups from one site and the group of healthy adults was highly significant (p less than 0.001). In some newborns we found also elevated percent-age of B lymphocytes with the receptor for C3 (EAC test) on 5-th day of life. The difference between this group and adults was not so great (0,02 less than p less than or equal to 0.05). It was also stated that absolute counts of B lymphocytes estimated by EA and EAC tests was elevated in children below 10 years of age.

In order to obtain higher concentration of IgA-antibodies in commercial gammaglobulin preparations, used for treatment of hipo - and dysimmunoglobulinemia states, there are produced two preparations: "Gamma-A- Konzentrat " of Behringwerke and " Igalina " of Biomed . The first one was examined and described by us previously (4, 12). Now we present our studies on Igalina . The examinations were performed using electrophoresis on cellulose acetate, double diffusion in agar gell using 15 specific antisera to plasma proteins and immunoelectrophoresis with anti-IgG, IgA and IgM. There were detected traces of albumin, transferin , alfa2 macroglobulin, haptoglobin fibrinogen and presence of IgA, IgG and IgM. We also observed the split of IgG line in immunoelectrophoresis at the cathode and to Fc and Fab fragments. The concentrations of IgA, IgG and IgM were evaluated using Mancini method (IgA--920 IU/ml, IgG--1640 IU/ml IgM 2500 IU/ml). Concentrations of chosen antibodies in Igalina were: ASO--1000 U/ml, anti- Staphylolisins 8 U/ml, agglutinins antistaphilococal 1 : 320, diphtheria antitoxins 42 U/ml, agglutinins antithyphi 1 : 20----1 : 640. They were 4 to 100 times higher than in plasma serum. Isohaemagglutinins were not detected. Igalina was used for preventive treatment of children contacted with viral infections or therapeutically for children who demonstrated first symptoms of viral infections of respiratory tracts. Preparation was given 48 times to 40 children. This group included 19 newborns and 10 children with I and IV type of dysimmunoglobulinemia or hipoimmunoglobulinemia . Doses ranged from 0,5 ml up to 1,0 ml/kg body weight. Estimations of IgA, IgG and IgM concentrations in serum and saliva of children (using Mancini method) were carried out before injection of Igalina and on the third and seventh day after injection. Results of studies were statistically evaluated, and besides the lack of significant differences we found some rise of IgA and IgG concentrations. We would like to underline also that in 6 children with hipo or dysimmunoglobulinemia the levels of IgA and IgG reached normal value on the third day after injection of Igalina . Investigations of secretory piece in saliva in the same period were carried out too. In three patients we could find the presence of secretory piece just after injection of Igalina . We did not found antibodies to IgA in patients' sera after treatment. The clinical results of treatment as well as toleration of the drug were good and very good.

The study has aimed at investigating the blastic transformation of peripheral blood lymphocytes in patients with pituitary dwarfism before, during and after the treatment with human growth hormone. The reactivity of lymphocytes in vitro was studied in cultures stimulated with phytohemagglutinin-M of the Difco Company (PHA), in cultures stimulated with human growth hormone (HGH), and in cultures non-stimulated in vitro. The investigations were carried out in 34 patients with pituitary dwarfism at the age of 7 to 19 years. The control group was made up of 16 healthy children, 8 to 15 years old who had never been administered HGH. The patients were divided into groups according to the circumstance whether it was the first or the subsequent contact with HGH, and according to the periods at which the blood samples were collected during after therapy. It was found that the capability of lymphocytes to blastic transformation after stimulation with a specific stimulus (HGH) in patients with pituitary dwarfism in comparison with the reaction of healthy children is reduced (Fig. 2). The ability of the transformation of lymphocytes in this case shows selectivity towards the stimulus because the inhibiting of the response was noted only after the stimulation with HGH, but not after stimulation with PHA. During the treatment with HGH, allergization of the body appears; this is evidenced by the increase in the percentage of blastic cells in autotransformation (Fig. 1).

The effect of hydrocortisone on immunologically competent cells in peripheral blood (lymphocytes B and RFC cells) and on the level of humoral antibodies has been studied in rabbits immunized twice with 10per cent sheep erythrocytes. The animals were divided into two groups. The experimental group, numbering 14 rabbits, was administered 30 mg/kg body weight of hydrocortisone intramuscularly for three consecutive days before the first and second dose of antigen; the 10 rabbits of the control group were administered only the antigen. The results obtained showed that, after immunization and in the course of the normal immune response of the body, the numbers of RFC cells and of lymphocytes B increased in peripheral blood before the level of antibodies rose (Fig. 1, 3 and 4). The results in the group of experimental rabbits showed that hydrocortisone markedly suppressed the normal immune response of the body to the antigen administered. When the data obtained in both groups studied were compared, it was found that the amount of RFC cells and of lymphocytes B in peripheral blood was manifestly smaller in the group of experimental animals treated with hydrocortisone, than in the group of control animals, both after the first and the second dose of antigen (Fig. 1, 2 and 3). The difference concerning the humoral antibodies level between the two groups, however, appeared only after the second dose of the antigen (Fig. 4, Table I). That means that the investigation of immunologically competent cells in peripheral blood, such as lymphocytes B and RFC cells, enables to detect disturbances in the immune response of the body earlier than the investigation of humoral antibodies and supplies a more complete image of the changes occurring in the body during immunization.