US2020232985A1PendingUtilityA1

IgG SUBTYPING ASSAY FOR IDENTIFYING TRANSPLANTABLE TISSUE SAMPLES

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Assignee: NEW JERSEY ORGAN AND TISSUE SHARING NETWORK INCPriority: Dec 3, 2015Filed: Apr 7, 2020Published: Jul 23, 2020
Est. expiryDec 3, 2035(~9.4 yrs left)· nominal 20-yr term from priority
Inventors:Prakash Rao
G01N 33/56972A61K 35/34G01N 33/6854A61K 35/22G01N 33/564G01N 15/14G01N 2015/1488
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Claims

Abstract

The disclosure relates to methods for assessing the suitability of a tissue obtained from a vertebrate (e.g., human) donor for grafting to a recipient, such as another vertebrate of the same species. The method involves contacting leukocytes obtained from the donor with serum from the recipient. Binding between recipient antibodies and the leukocytes is assessed, specifically focusing on the subtype(s) of IgG antibodies which bind with donor leukocytes. Detected binding between recipient antibodies of subtypes IgG1 and IgG3 indicates that the donor tissue is not suitable for grafting to the recipient. Detected binding between recipient antibodies of subtypes IgG2 and/or IgG4 indicates that the donor tissue can suitably be grafted to the recipient.

Claims

exact text as granted — not AI-modified
What is claimed is: 
     
         1 . A method of grafting a tissue obtained from a vertebrate donor to a recipient, the method comprising
 contacting leukocytes obtained from the donor with antibodies obtained from the recipient,   assessing binding between the donor leukocytes and recipient antibodies of at least IgG subtypes IgG1 and IgG3, and   grafting the donor tissue to the recipient if substantially no binding between the donor leukocytes and recipient antibodies of either of IgG subtypes IgG1 and IgG3 is detected.   
     
     
         2 . The method of  claim 1 , comprising assessing binding between the donor leukocytes and recipient antibodies of at least IgG subtypes IgG1, IgG2, IgG3, and IgG4. 
     
     
         3 . The method of  claim 2 , wherein assessment of binding between the donor leukocytes and recipient antibodies of each of IgG subtypes IgG1, IgG2, IgG3, and IgG4 is performed in a single assay. 
     
     
         4 . The method of  claim 1 , wherein the assessment of binding between donor leukocytes and the antibodies is performed by contacting recipient antibodies with differentially-labeled antibodies that specifically recognize antibodies of the IgG1 and IgG3 subclasses. 
     
     
         5 . The method of  claim 1 , wherein the assessment of binding between donor leukocytes and the antibodies is performed by contacting recipient antibodies with differentially-labeled antibodies that distinguishably recognize antibodies of the IgG1, IgG2, IgG3, and IgG4 subclasses. 
     
     
         6 . The method of  claim 5 , further comprising contacting the donor leukocytes with differentially-labeled antibodies that distinguishably recognize B and T cells of the donor. 
     
     
         7 . The method of  claim 1 , wherein individual donor leukocytes are assessed by flow cytometry after contacting the donor leukocytes with the recipient antibodies. 
     
     
         8 . The method of  claim 1 , wherein individual donor leukocytes are assessed by flow cytometry after contacting the donor leukocytes with the recipient antibodies and after contacting recipient antibodies with differentially-labeled antibodies that distinguishably recognize antibodies of the IgG1, IgG2, IgG3, and IgG4 subclasses. 
     
     
         9 . The method of  claim 1 , wherein individual donor leukocytes are assessed by flow cytometry after contacting the donor leukocytes with the recipient antibodies, after contacting recipient antibodies with differentially-labeled antibodies that distinguishably recognize antibodies of the IgG1, IgG2, IgG3, and IgG4 subclasses, and after contacting the donor leukocytes with differentially-labeled antibodies that distinguishably recognize B and T cells of the donor. 
     
     
         10 . The method of  claim 1 , wherein the recipient is a vertebrate. 
     
     
         11 . The method of  claim 10 , wherein the recipient is the same species as the donor. 
     
     
         12 . The method of  claim 11 , wherein the recipient is a human. 
     
     
         13 . The method of  claim 1 , wherein non-specific binding between recipient antibodies and Fc receptors on donor leukocytes is inhibited. 
     
     
         14 . The method of  claim 13 , wherein the non-specific binding is inhibited by contacting the donor leukocytes with a peptide that inhibits binding between antibodies and surface Fc receptors prior to contacting the donor leukocytes with the recipient antibodies. 
     
     
         15 . The method of  claim 1 , wherein the donor leukocytes are alive at the time they are contacted with the recipient antibodies. 
     
     
         16 . A method of assessing compatibility of a body tissue of a potential donor for grafting with a recipient, the method comprising
 contacting leukocytes obtained from the potential donor with antibodies obtained from the recipient,   assessing binding between the potential donor leukocytes and recipient antibodies of at least IgG subtypes IgG1 and IgG3,   whereby the body tissue is assessed as compatible for grafting with the recipient if substantially no binding between the donor leukocytes and recipient antibodies of either of IgG subtypes IgG1 and IgG3 is detected.   
     
     
         17 . The method of  claim 16 , comprising assessing binding between the donor leukocytes and recipient antibodies of at least IgG subtypes IgG1, IgG2, IgG3, and IgG4. 
     
     
         18 . The method of  claim 17 , wherein assessment of binding between the donor leukocytes and recipient antibodies of each of IgG subtypes IgG1, IgG2, IgG3, and IgG4 is performed in a single assay. 
     
     
         19 . The method of  claim 16 , wherein the assessment of binding between donor leukocytes and the antibodies is performed by contacting recipient antibodies with differentially-labeled antibodies that specifically recognize antibodies of the IgG1 and IgG3 subclasses. 
     
     
         20 . The method of  claim 16 , wherein the assessment of binding between donor leukocytes and the antibodies is performed by contacting recipient antibodies with differentially-labeled antibodies that distinguishably recognize antibodies of the IgG1, IgG2, IgG3, and IgG4 subclasses. 
     
     
         21 . The method of  claim 20 , further comprising contacting the donor leukocytes with differentially-labeled antibodies that distinguishably recognize B and T cells of the donor. 
     
     
         22 . The method of  claim 16 , wherein individual donor leukocytes are assessed by flow cytometry after contacting the donor leukocytes with the recipient antibodies. 
     
     
         23 . The method of  claim 16 , wherein individual donor leukocytes are assessed by flow cytometry after contacting the donor leukocytes with the recipient serum and after contacting recipient antibodies with differentially-labeled antibodies that distinguishably recognize antibodies of the IgG1, IgG2, IgG3, and IgG4 subclasses. 
     
     
         24 . The method of  claim 16 , wherein individual donor leukocytes are assessed by flow cytometry after contacting the donor leukocytes with the recipient serum, after contacting recipient antibodies with differentially-labeled antibodies that distinguishably recognize antibodies of the IgG1, IgG2, IgG3, and IgG4 subclasses, and after contacting the donor leukocytes with differentially-labeled antibodies that distinguishably recognize B and T cells of the donor. 
     
     
         25 . The method of  claim 16 , wherein the recipient is a vertebrate. 
     
     
         26 . The method of  claim 25 , wherein the recipient is the same species as the donor. 
     
     
         27 . The method of  claim 26 , wherein the recipient is a human. 
     
     
         28 . The method of  claim 16 , wherein non-specific binding between recipient antibodies and Fc receptors on donor leukocytes is inhibited. 
     
     
         29 . The method of  claim 28 , wherein the non-specific binding is inhibited by contacting the donor leukocytes with a peptide that inhibits binding between antibodies and surface Fc receptors prior to contacting the donor leukocytes with the antibodies obtained from the recipient. 
     
     
         30 . The method of  claim 1 , wherein the donor leukocytes are alive at the time they are contacted with the antibodies obtained from the recipient.

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