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Biomedical subjects

S N Saxena

Publications and source records attributed to S N Saxena.

At least 37 records · Page 2Linked to original sources

Effect of different concentrations of diphtheria toxoid & aluminum phosphate on potency of diphtheria component in DPT vaccines.

An effort was made to determine the optimum concentration of diphtheria toxoid in combination with aluminum phosphate gel in DPT vaccines which may give a safe, potent and economical preparation. The effect of four different concentrations of aluminum phosphate and three different antigenic concentrations of diphtheria toxoid on potency of diphtheria component in DPT vaccine was assessed. A gradual increase in potency was seen with increase in toxoid concentration and a gradual decrease in potency with the increase in aluminum phosphate content. Vaccines made with minimum quantities of toxoid (30 Lf/ml) and aluminum phosphate (3 mg/ml) were found to be highly satisfactory. Vaccines prepared with high antigenic purity toxoid have better potency, as compared to those prepared with a relatively low antigenic purity toxoid.

Adjuvants, Immunologic↗

Indirect haemagglutination test for the assay of antibodies against Japanese encephalitis viral antigen in human & animal sera.

The indirect haemagglutination (IHA) test was standardized for the assay of antibodies against Japanese encephalitis (JE) virus. Glutaraldehyde fixed sheep erythrocytes were sensitized with purified and concentrated JE vaccine (200-300% brain concentration). The JE vaccine made from Nakayama-NIH strain of JE virus was purified by protamine sulphate treatment and by ultracentrifugation in a sucrose gradient. The sensitized cells were quite stable in liquid as well as in lyophilized state both at -70 degrees C and 4-8 degrees C. These cells could be used for two years without much loss (4-8 times loss) in titre. The IHA test was as sensitive as the neutralization (N) test performed by plaque reduction method in chick embryo fibroblasts. The sensitivity of the test was influenced by the source of erythrocytes i.e., from the different sheep from which these were drawn. After standardization of the test, 16 human sera and 18 sera of immunized mice were assayed for antibodies against JE virus by N and IHA tests. There were no significant differences between titres of both human and mice sera determined by N and IHA tests (P greater than 0.05). The correlation coefficient between N and IHA titres for human sera was 0.60 (P less than 0.05) and for mice sera 0.82 (P less than 0.01). The IHA test has been found to be very simple, inexpensive, sensitive and reproducible.

Animals↗

Correlation between mouse intracerebral potency & 50 per cent histamine sensitizing dose of pertussis vaccine.

A good correlation was observed between mouse intracerebral potency in terms of International Units and 50 per cent histamine sensitizing dose (HSD50) of various types of pertussis vaccine. The correlation was significant irrespective of the inactivating agent used for the preparation of pertussis vaccine. If the potency of the vaccine increased, the HSD50 decreased and vice versa showing a negative correlation between potency and HSD50. The relative histamine sensitizing (HS) activities of various types of pertussis vaccines in comparison to that of heat inactivated pertussis vaccine (HIPV) obtained in different experiments were very similar although the HSD50 values of same types of vaccines obtained in these experiments varied greatly. The conventional HIPV preparations having potency around 8 IU per 40 x 10(9) organisms had HSD50 values in the range of 1.0 to 2.2 x 10(8) organisms. The estimation of HSD50 of pertussis vaccine has been found to be a useful measure of the toxicity of the vaccine and also correlates well with the potency of the vaccine.

Animals↗

Humoral and cell mediated immune responses to porins of Salmonella typhi.

Humoral and cell mediated immune responses were studied in control, infected and immunized-infected mice at different time intervals. The levels of antiporin antibodies were found to be higher throughout the study period in immunized-infected group in which 87.5% protection was observed by mouse potency test. A significant increase in stimulation index of lymphocyte blast transformation as well as delayed type hypersensitivity response was observed in the same group as compared to infected and control group when porins were used as antigens. Using nonspecific mitogens like PHA and Con A the lymphocyte proliferation was maximum in control group whereas a significant depression was observed in infected mice. It is indicated from this study that porins are excellent antigens interacting efficiently with both arms of the host immune systems which could play a role in providing protection against the disease.

Animals↗

Immune status of volunteers one year after administration of Japanese encephalitis vaccine produced in India.

The immune status of 40 volunteers who received the full course of Japanese encephalitis (JE) vaccine a year earlier and 15 individuals who had received only a booster dose at the same time, was studied by estimating the level of persistence of protective antibody in the sera. All the sera showed persistence of 100 per cent seroconversion rate. Individuals who had the full course of vaccination still had high levels of antibody (mean 2.8 Iog10); however there was a fall of 0.8 Iog10 from the post-booster level. Volunteers who had received only a booster dose, also showed persistence of high level of protective antibody (mean 2.4 Iog10), a drop of 0.9 Iog10 from the post-booster level. Neutralizing (N) antibody estimated using Dibrugarh (7812474) strain of JE virus also demonstrated persistence of high level of protective antibody against this virus (mean 2.4 Iog10). Persistence of high level of protective antibody against homologus and heterologus (Dibrugarh) virus strains and absence of vaccine related side-effects even one year after administration of JE vaccine produced in India, demonstrates the immunizing potency and safety of this new vaccine.

Adolescent↗

Surveillance of salmonellae in India for drug resistance.

Out of 3001 salmonella received during 1984-85, 2985 strains were tested for antibiotic resistance and R-pattern transfer. Multidrug resistance was observed in 73% of diarrhoeal and 43% of non-diarrhoeal isolates. Antibiotic resistance markers were transferable in 46.6% of diarrhoeal and 41.1% of non-diarrhoeal isolates. Two out of 4 salmonella sero-types isolated for the first time in India during this period were multidrug resistant.

Anti-Bacterial Agents↗

Evaluation of the new rapid rabies immunodiagnosis technique.

The rapid rabies enzyme immuno-diagnosis (RREID) kit was evaluated for its sensitivity and specificity in comparison to fluorescent antibody technique (FAT) and biological test (BT) for the diagnosis of rabies. While 93 per cent correlation was observed in case of fresh brain samples, 71 per cent correlation was noted with glycerol preserved samples, where RREID was more sensitive than FAT. RREID test can be employed for rapid diagnosis of rabies, when facilities for FAT are not available. However, to test, at one time, small number of specimens, the RREID kit needs to be modified.

Animals↗

Development of a mouse model to estimate the potency of the diphtheria toxoid component of diphtheria-tetanus and diphtheria-tetanus-pertussis vaccines.

A mouse model to estimate the potency of the diphtheria toxoid component in diphtheria-tetanus vaccines and diphtheria-tetanus-pertussis vaccines has been developed as an alternative to the conventional method of testing in guinea-pigs. Optimal conditions with regard to dose, route and period of immunization have been standardized. The maximum levels of antitoxin were detected five weeks after vaccination and the s.c. route was found to be optimal. Potency data have been compared with other studies in mouse models and with those obtained by the conventional method in guinea-pigs.

Animals↗

The effects of purified pertussis components and lipopolysaccharide on the results of the mouse weight gain test.

The effects of highly purified components of Bordetella pertussis, that is pertussis toxin (PT) and filamentous haemagglutinin (FHA), and of lipopolysaccharide (LPS) were studied in the active mouse weight gain test (MWGT). The PT when given alone or with other components in various combinations caused weight losses and deaths 2-3 days after inoculation but FHA was not toxic in the MWGT. When FHA was given with PT, the toxic effect of PT was reduced. The LPS caused weight losses at 24 h which decreased when LPS was given with PT. The toxic effects of PT as indicated by late deaths and late weight losses or failure to gain weight continued until 14 days after inoculation. The various components had similar effects on mouse weight gain in both LACA and NIH strains of mice. The doses of PT used in the MWGT caused marked leucocytosis but FHA and LPS did not. No agglutinins appeared in the sera of mice inoculated with various purified components. The components were thus pure and did not contain agglutinogens.

Agglutinins↗

The reduction of animal usage by the application of indirect haemagglutination in the potency testing of diphtheria toxoid in combined vaccines.

Eight Adsorbed Diphtheria-Tetanus vaccines and 13 Diphtheria-Tetanus-Pertussis vaccines made by four different manufacturers were tested for the potency of the diphtheria components in guinea-pigs by the method of British Pharmacopoeia (1973). Two-hundred-and-ten guinea-pig sera consisting of ten sera related to each vaccine sample thus obtained were titrated for diphtheria antitoxin by indirect haemagglutination (IHA) and the conventional toxin neutralization (TN) tests. Statistical analysis of the results showed a good correlation between the titres obtained with the two tests. The potencies of the diphtheria components of various vaccines calculated from the antitoxin content of the respective guinea-pig sera titrated by the IHA test correlated significantly with the potencies obtained from the antitoxin content titrated by the routinely used TN test. The use of IHA in place of the TN test thus offers as an alternative that permits a reduction in animal usage.

Animal Testing Alternatives↗

Studies on the optimal conditions for inactivation of Bordetella pertussis organisms with glutaraldehyde for preparation of a safe and potent pertussis vaccine.

The optimal conditions for inactivation of Bordetella pertussis organisms with glutaraldehyde for the production of a safe and potent whole cell pertussis vaccine were investigated. Two bacterial harvests from B. pertussis strain 10536 were treated with glutaraldehyde, each with 0.025, 0.05 and 0.1% concentrations of glutaraldehyde for 10, 60 and 120 min. The nine types of glutaraldehyde-inactivated pertussis vaccine (GIPV) and conventional heat-inactivated pertussis vaccine (HIPV) preparations made from two bacterial harvests were comparatively evaluated for the mouse weight gain test (MWGT), potency, and the histamine-sensitization (HS) and leucocytosis-promoting-factor (LPF) tests. The minimum period for killing the B. pertussis organisms with glutaraldehyde was greater than 10 min for 0.025%, 10 min for 0.05% and 5 min for 0.1% concentration. The average loss in opacity varied from 5 to 10% for GIPV preparations and was 14% for HIPV preparations. The GIPV preparations except those inactivated with 0.025% glutaraldehyde for 10 min (GIPV-A) were much less toxic than the HIPV preparations in the MWGT. The GIPV-A preparations did not pass the MWGT. The GIPV preparations were also much less toxic in HS and LPF tests than the HIPV preparation. The potency of GIPV preparations inactivated with 0.05% glutaraldehyde for 10 min (GIPV-D) was similar to that of HIPV preparations. The prolonged treatments with glutaraldehyde reduced the potency. The GIPV-D preparation with good potency and less toxicity was found to be inactivated with glutaraldehyde under optimal conditions. All the preparations were innocuous in the abnormal toxicity test.

Aldehydes↗

The effects of different inactivating agents on the potency, toxicity and stability of pertussis vaccine.

The effect of heat (56 degrees C for 10 min), formaldehyde (0.1% at 37 degrees for 24h), glutaraldehyde (0.05% at room temperature for 10 min), thimerosal (0.02% at 37 degrees C for 24h), acetone-I (three treatments at room temperature) and acetone-II (three treatments at room temperature and fourth treatment at 37 degrees C), when used as inactivating agents in the preparation of pertussis suspension, was studied with regard to potency, toxicity and stability. Five batches each of Bordetella pertussis strains 134 and 509 were used for the study. The thimerosal inactivated pertussis (TIP) preparation was 1.5-2 times more potent than the heat inactivated pertussis (HIP) preparation. The potency values of the formaldehyde inactivated pertussis (TIP) and glutaraldehyde inactivated pertussis (GIP) preparations were similar to those of the HIP preparation, while the potencies of the acetone-I treated pertussis (A(I)TP) and acetone-II treated pertussis (A(II)TP) preparations were about half those of the HIP preparation. The FIP preparation was the least toxic showing maximum weight gain in the mouse weight-gain test (MGWT), while the TIP preparation did not pass the MWGT. The weight gains shown the GIP, A(I)TP and A(II)TP preparations were greater than those shown by the HIP preparation. The potency of pertussis component in the adsorbed diphtheria-pertussis-tetanus (DPT) vaccine was stable at 4-8 degrees C and 25 degrees C for three months for all types of pertussis vaccine. There was about 54-65% loss in the potency of the samples after three months at 35 degrees C. The inactivating agents used in the manufacture of pertussis preparations had no effect on the stability of the vaccine.

Acetone↗