An enzyme-linked immunosorbent assay of anti-sheep red blood cell antibodies of the classes M, G, and A in the rat

Loveren, Henk Van; Verlaan, A. P. J.; Vos, J.G.

doi:10.1016/0192-0561(91)90181-6

Cited by 22 publications

(15 citation statements)

References 9 publications

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“…The levels of antibodies were evaluated after the stimulation of the immune system with SRBC and the administration of DMDE for thirty days (Figure 3). Experiments on immunization of animals with a suspension of intact sheep red blood cells in a standardized concentration are widely described in the literature (Delaney et al, 2001;Ladics et al, 1995, Temple et al, 1993Van Loveren et al, 1991), and some researchers have already used the suspension of industrialized SRBC (BioMérieux ® ) (Alvarez et al, 2004).…”

Section: Discussionmentioning

confidence: 99%

“…Three days prior to sacrificing the animals, a reinforcement dose of 50 µg/500 µL of SRBC soluble antigen was administered intravenously (i.v.) (Ladics et al, 1995;Temple et al, 1993;Van Loveren et al, 1991 After thirty days, serum samples from these animals were collected, and the animals were sacrificed by decapitation. After aseptic removal, each spleen was evaluated macroscopically, and approximately one-third of each spleen was separated for histological studies.…”

Section: Immunization Of the Animalsmentioning

confidence: 99%

“…The SRBC antigen were determined by the method described by Temple et al, 1993;Van Loveren et al,1991.…”

Section: Membrane Antigen From Sheep Red Blood Cells (Srbc)mentioning

confidence: 99%

“…After developing, the absorbancies were read at 492 nm with an automatic reader for microplates (Arriba et al, 2002;Ladics et al, 1995;Temple et al, 1993;Van Loveren et al, 1991).…”

Section: Elisa Measurement Of the Igg Response To Srbcmentioning

confidence: 99%

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Evaluation of the immunotoxicological effects of Dimorphandra mollis Benth., Fabaceae, in rats

Feres¹,

Toledo²,

Tagliati³

et al. 2010

Rev. bras. farmacogn.

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RESUMO: "Avaliação dos efetos imunotoxicológicos de Dimorphandra mollis Benth., Fabaceae, em ratos". A Dimorphandra mollis Benth., Fabaceae, conhecida como faveira ou favad´anta, é uma planta comum do cerrado central do Brasil, muito utilizada por suas propriedades antioxidante, antiplaquetária e, principalmente, como vasoprotetora. Seu principal marcador é a rutina. Este estudo teve como objetivo avaliar a segurança da utilização do extrato seco de D. mollis em roedores. O extrato foi extraído, padronizado e quantificado apresentando teor de 76,0±3% de rutina. Nos parâmetros bioquímicos e hematológicos avaliados, não se observou alterações nos grupos de machos e fêmeas que receberam a dose de 1000 e 2000 mg/kg de D. mollis, mas observou-se um aumento de eosinófilos. Nos estudos histopatológicos detectou-se hiperreatividade da polpa branca esplênica, no grupo que recebeu a dose de 2000 mg/kg de D. mollis. Na avaliação da resposta linfoproliferativa, com 1000 e 2000 mg/kg não foram observadas alterações, e somente nos estudos com a dose de 2000 mg/kg se observou diminuição de IgG. Os resultados obtidos, utilizando roedores, sugerem que nenhuma toxicidade existe na administração de extrato seco de D. mollis na dose de 1000 mg/kg. Unitermos: Dimorphandra mollis, rutina, imunotoxicologia, roedores.ABSTRACT: Dimorphandra mollis Benth., Fabaceae, also known as "faveira" or "fava-d´anta", is a plant common to the central woodsy meadow region of Brazil. It is well known for its antioxidant, antiplatelet and, principally, vasoprotective properties. Its principal component is rutin. The objective of this study is the evaluation of the safety of the use of the dried D. mollis extract in rodents. The rutin content of the standardized extract was 76.0±3%. With respect to the biochemical and hematological parameters evaluated, no alterations in the groups of rats that received 1000 and 2000 mg/kg doses of D. mollis were observed, but an increase in eosinophiles occurred. Hyperactivity of the white splenic pulp was detected in the group that received the 2000 mg/kg dose of D. mollis. In the evaluation of the lymphproliferative response with 1000 and 2000 mg/kg, no alterations were observed, and a decrease in IgG was only observed in the studies with a 2000 mg/kg dose. The results obtained with rodents suggest that no toxicity exists with the administration of dried D. mollis extract in a 1000 mg/kg dose.

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Section: Discussionmentioning

confidence: 99%

Section: Immunization Of the Animalsmentioning

confidence: 99%

“…The SRBC antigen were determined by the method described by Temple et al, 1993;Van Loveren et al,1991.…”

Section: Membrane Antigen From Sheep Red Blood Cells (Srbc)mentioning

confidence: 99%

“…After developing, the absorbancies were read at 492 nm with an automatic reader for microplates (Arriba et al, 2002;Ladics et al, 1995;Temple et al, 1993;Van Loveren et al, 1991).…”

Section: Elisa Measurement Of the Igg Response To Srbcmentioning

confidence: 99%

See 2 more Smart Citations

Evaluation of the immunotoxicological effects of Dimorphandra mollis Benth., Fabaceae, in rats

Feres¹,

Toledo²,

Tagliati³

et al. 2010

Rev. bras. farmacogn.

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“…Enzyme-linked immunosorbent assay (ELISA) methods using whole SRBC (Heyman et al, 1984;Mori et al, 1989) and SRBC membranes (Van Loveren et al, 1991;Temple et al, 1993) have been reported. Because of the long-term stability of the membranes, an SRBC-ELISA using the cell membranes has been extensively used to detect T-lymphocyte-dependent antigen-induced primary antibody responses (Temple et al, 1995).…”

Section: Introductionmentioning

confidence: 99%

Use of Whole Sheep Red Blood Cells in ELISA to Assess ImmunosuppressionIn Vivo

Koganei¹,

Tsuchiya²,

Samura³

et al. 2007

Journal of Immunotoxicology

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An enzyme-linked immunosorbent assay (ELISA) using whole sheep red blood cells (SRBC) has been reported as one of the methods for detecting a T-lymphocyte-dependent antibody response. However, it has not been widely used because of SRBC problems such as the weak attachment to ELISA plates, specificity and shortterm stability. The objectives of this study were to address these issues and to validate the SRBC-specific antibody response assay. Male Sprague-Dawley rats were bled after 6 days of SRBC immunization. In our new procedure, glutaraldehyde was added before discarding the supernatant of inoculated SRBC suspension to attach SRBC firmly to the plate, while in the original method it was added after discarding. As a result, the attached SRBC was maintained throughout the ELISA procedures. No interference was observed in the titration curve of IgM and IgG antibodies in rats and IgM-antibody in mice when control sera were analyzed to evaluate specificity of this method. The short-term stability of SRBC was overcome by using the different lots of SRBC. They provided antibody titers, which were consistent with those measured using the same lot for immunization. In addition, cyclophosphamide, cyclosporine, prednisolone and methotrexate, well-known immunosuppressive agents, were tested to confirm the applicability of the improved ELISA method to detect the T-lymphocyte-dependent antibody response. All four compounds inhibited the IgM antibody responses dose-dependently. These results demonstrate that the improved whole SRBC-ELISA method provides reproducible and reliable results in the T-lymphocyte-dependent antibody response assay.

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