Tetanus Assay

Tetanus (adsorbed) Cultures of Cl. tetani in liquid medium 1 Conversion of toxin to toxoid 2 Separation and purification of toxoid 3 Adsorption to adjuvant 3 + 3 quantal assay in mice using subcutaneous challenge with tetanus toxin Inoculation of guinea-pigs to exclude presence of untoxoided toxin... 

In rodents, sheep red blood cells (SRBC) are routinely used for immunization. The antibody response is determined using the plaque forming cells assay (PFC) or by plasma SRBC-specific antibody titer [21, 22], As an alternative to SRBC, other T-cell-dependent antigen may be used, including keyhole limpet hemocyanin (KLH), tetanus toxoid (TT), or pneumococcal antigen. 

The delayed type hypersensitivity response (DTH) is an assay frequently used to assess the T cell response to commonly encountered microbial antigens. It involves intradermal injection of antigens to which the majority of individuals are immune (known as recall antigens) such as vaccinia, herpes simplex, and mumps viruses, Candida, and tetanus toxoid. In normal individuals, after 24-48 hours, an inflammatory filtrate results in local edema and induration, the diameter of which can be measured. A negative reaction to all the antigens (anergy) is usually reflected by decreased lymphocyte function as measured in vitro and is frequently seen in AIDS and ARC patients. 

AES is used in pharmacopoeial assays of (1) Na in albumin solution and plasma protein solution (2) K, Na and barium (Ba) in calcium acetate used to prepare dialysis solutions (3) Ca in adsorbed vaccines (e.g. diphtheria and tetanus). 

A six-year-old child had anaphylaxis 30 minutes after a fifth dose of DT vaccine (22). Skin tests, in vitro determination of specific IgE antibodies, and immunoblotting assays showed that the IgE response was directed against tetanus and diphtheria toxoids. Cross-reactivity between the two toxoids was not demonstrated, indicating the presence of co-existing but non-cross-reacting IgE and IgG antibodies. 

Sanchez-Prieto J, Shira TS, Evans D, Ashton A, Dolly JO, Nicholls DG (1987) Botulinum toxin A blocks glutamate exocytosis from guinea-pig cerebral cortical synaptosomes. In Eur. J. Biochem. 165 675-81 Schiavo G, Montecucco C (1995) Tetanus and botulism neurotoxins isolation and assay. Methods Enzymol. 248 643-52... 

Development of human in vitro systems will require optimization of stimulator (preferably using antigen relevant to human exposure, such as TT [tetanus toxin]), culture conditions, and assay endpoint(s). For these reasons, further research in this area is strongly recommended. 

Holmstrom et al. (1989b) evaluated the long-term inhalation effects of formaldehyde exposure to immune function in female Sprague-Dawley rats exposed to 12.6 ppm formaldehyde for 6 hours/day, 5 days/week for 22 months. After 22 months of formaldehyde exposure, the rats were inoculated subcutaneously with Pneumovax (Merck Sharpe and Dohme) and antitetanus vaccine (National Bacteriological Laboratory). Animals were sacrificed at 21-25 days after vaccination. Blood samples were collected from each animal before vaccination and just prior to sacrifice. Tlie blood was analyzed for response to Pneumovax and tetanus vaccination using an enzyme-linked immunosorbant assay technique. The results indicated... 

Effects on immune function, as indicated by altered responses in humoral and cell-mediated immunity assays and host resistance tests, were also induced by intermediate-duration oral exposure to commercial mixtures. Studies in nonprimate species showed reduced antibody responses to tetanus toxoid in guinea pigs exposed to Clopen A-60 (4 mg/kg/day for 3-5 weeks), keyhole limpet hemocyanin in rats exposed to Aroclor 1254 (4.3 mg/kg/day for 10 weeks), and SRBC in mice exposed to Aroclor 1242 (22 mg/kg/day... 

Evaluation of a T cell-dependent antibody response (TDAR) is a critical component of the evaluation of immune function in the neonates. In this assay, neonates are injected with a T-dependent antigen, usually keyhole hmpet hemocyanin (KLH) or tetanus toxoid ( ITX), and the development of antibodies is measured over time. In monkeys there can be high inter-animal variability so the collection of serial blood samples is important and the data can be expressed as the area under the antibody titer versus time curve. Figure... 

Figure 6 Proliferation of T cells from mice immunized with nanoparticulate of soluble formulations in response to ex vivo stimulation with the recall antigen (tetanus toxoid). The T cells were derived from the mice immunized with TT and various doses of CpG ODN. The number of T cells and APCs incubated in the assay plate wells were 5x10 and 1x10 respectively. The recall proliferation response is shown on the y-axis as (A) CPM for clarity SI values are also given on the top of each bar (B) stimulation index for nanoparticulate (solid circles) and soluble (hollow circles) mode of vaccine delivery. Error bars indicate standard deviation. (From Ref. 135.)...

Acellular pertussis antibody response ELISA assays [13] were run in the study of the acellular pertussis vaccine component of the Diphtheria and Tetanus Toxoids and acellular Pertussis Vaccine Adsorbed (DTaP) to assess vaccine potency before and after irradiation of DTaP Vaccine Lots A, B, and C. 

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