Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Reexamination Certificate
1996-09-10
2001-07-31
Caputa, Anthony C. (Department: 1642)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
C435S007200, C435S007210, C435S007240, C435S007920, C435S007940
Reexamination Certificate
active
06268155
ABSTRACT:
BACKGROUND OF THE INVENTION
The assessment of inflammatory responses has been achieved by subjective analysis either by actual measurement and scoring or through the use of immunohistochemistry of the inflammatory response. Among the types of inflammatory responses is delayed type hypersensitivity response (DTH), which results in redness and swelling, due to exposure to a sensitizing antigen, similar to an allergic reaction.
In order to objectively quantify cellular infiltration in a skin DTH response a novel methodology was developed. The procedure involves a new method for solubilizing a skin biopsy and then quantifying the surface antigen expressed by the infiltrating leukocytes in the solubilized skin biopsy. This quantification is made possible by applying a capture ELISA (enzyme linked i mnmunoadsorbant assay) using two monoclonal antibodies. This methodology will allow for the quantification of DTH responses, as well as any cellular antigens in tissues and biopsies, provided there are two monoclonal or polyclonal antibodies available to detect the antigen. An objective method for quantifying an inflammatory response to date has not been developed.
SUMMARY OF THE INVENTION
The instant invention relates to a method for quantitating a cellular infiltrate in a tissue biopsy of a warm-blooded animal comprising the steps of: (a) solubilizing the tissue biopsy to isolate a tissue antigen in the supernatant; and (b) measuring the surface antigen expressed by the cellular infiltration found in the tissue supernatant isolated from the tissue biopsy of the inflammatory response, using a capture ELISA method. This method also allows one to quantitate the cellular infiltration of a tissue biopsy corresponding to an inflammatory response. As such this invention has both research and clinical applications in the analysis of an inflammatory response, and the effectiveness of immunosuppressants in addressing the inflammation.
REFERENCES:
patent: 5202230 (1993-04-01), Kamentsky
patent: 5426029 (1995-06-01), Rittershaus et al.
Di Carlo, et al.,J. Neurosurg. Sci., vol. 34, No. 3-4, p. 181-5, Jul.-Dec. 1990. Abstract only.*
Kang, K. et al., “CD11b+ Macrophages That Infiltrate Human Epidermis After In Vivo Ultraviolet Exposure Potently Produce IL-10 and Represent the Major Secretory Source of Epidermal IL-10 Protein1”, J. Immunol., vol. 153, pp. 5256-5264, 1994.
Koo Gloria C.
Nguyen Mai P.
Talento Althea D.
Camara Valerie J.
Caputa Anthony C.
Daniel Mark R.
Merck & Co. , Inc.
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