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Laboratory for Immunologic Investigations

The Laboratory for Immunologic Investigations (L-II) is a novel endeavor at the Virginia-Maryland Regional College of Veterinary Medicine. The objective of the Laboratory is to define, develop, and standardize a battery of methods and protocols for use in evaluation of immunological competence of normal and diseased horses and other domestic species. The methods and protocols developed at the Laboratory are used to assess immune response, in both the clinical and research environments. Specifically, these techniques are used to characterize diseases in individual animals; evaluate, monitor, and predict species response to therapy; and further our general understanding of the immune regulation. Current activities of the L-II include establishment of immunological profiles for predominantly horses, although some effort has been made to accomplish the same task in other species of domestic animals.

The laboratory incorporates an interdisciplinary team of specialists, including veterinary clinicians board certified in internal medicine, nutrition, and epidemiology; along with basic immunologists. Faculty and laboratory specialists have expertise in immunologic and biochemical techniques, including tissue culture, cell isolation, cryopreservation, immunohistochemistry, and molecular biology. Some contributing investigators also have expertise in areas of specific equine diseases, like Equine Protozoal Myeloencephalitis, and Recurrent Airway Disease. Our objective is to develop standard operating procedures and good laboratory practice (GLP) for studies in immune investigation, and to apply these technologies to better understand immune response, especially in the diseased animal.

A. Instrumentation

Much of the instrumentation utilized by the laboratory is under the auspices of the BPRL facility including:

  • 4-color Coulter flow cytometer
  • Microplate reader
  • High speed centrifuges
  • Tissue culture lab
  • Radioisotope lab
  • Molecular biology lab

B. Technical Expertise

  • Cell isolation techniques, including lymphocyte, neutrophil, and monocyte isolation in equine, bovine, canine, and feline.
  • B lymphocyte evaluation, including immunoglobulin quantitation (IgG, M, A) and mono-clonal antibody techniques.
  • T lymphocyte evaluation, including monoclonal antibodies: CD4, CD8, CD3, CD11, CD18.
  • Lymphocyte blastogenesis/proliferation assays.
  • PCR analysis of equine cytokine mRNA.
  • Analysis of equine gene expression using macro array technology

C. Using the Laboratory for Immunologic Investigations

Persons interested in using the Laboratory for Immunologic Investigations should contact Dr. Virginia Buechner-Maxwell, the Laboratory Coordinator, at 1-3758.

D. Projects Currently in Progress

  • The role of lymphocytes apoptosis in equine recurrent airway obstruction.
  • The distribution and detection of maternal immunoglobulins into foal cerebrospinal fluid.
  • Effect of long term corticosteroid therapy on inflammatory cytokine expression in equine peripheral blood mononuclear cells.
  • The effect of dexamethasone therapy on equine peripheral blood immune cell function.
  • Evaluation of intradermal skin test in the horse.
  • Use of aerosolized magnesium as a treatment for recurrent airway obstruction in horses.
  • The affect of intense training on equine peripheral blood lymphocyte cytokine mRNA expression.
  • Altered surfactant composition and function in horses with recurrent airway obstruction.

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