Antibodies Against Cells of Fetal Origin

Abstract

This invention relates to antibodies that specific bind to fetal CD36+ cells in preference to binding to maternal CD36+ cells and methods for using these antibodies to detect and separate fetal cells from adult biological fluids including maternal peripheral blood.

1 - 23 . (canceled) 
     
     
         24 . A method for isolating CD36+ fetal cells from a biological fluid comprising the steps of:
 (a) isolating CD34+ cells from the biological fluid;   (b) stimulating the isolated CD34+ cells to express CD36;   (c) separating fetal cells from the non-fetal cells present in the isolated cells by contacting the isolated cells with one or a plurality of antibodies that specifically bind to a cellular antigen that is detectably expressed by CD36+ fetal cells, but not detectably expressed by CD36+ non-fetal cells; and   (d) isolating CD36+ fetal cells from step (c).   
     
     
         25 . The method of  claim 24 , wherein the antibody specifically binds to a cellular antigen identified as MS4A10, MS4A7, MS4A6A, ASGR2 or MS4A5. 
     
     
         26 . The method of  claim 25 , wherein the antibody specifically binds to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10. 
     
     
         27 . The method of  claim 26 , wherein the antibody specifically binds to SEQ ID NO:1. 
     
     
         28 . The method of  claim 24 , wherein the CD34+ cells are incubated in the presence of the cytokines Flt-3 ligand, TPO (thrombopoietin), and SCF (stem cell factor). 
     
     
         29 . The method of  claim 24 , wherein the fetal cells are separated from the non-fetal cells by fluorescence activated cell sorting (FACS). 
     
     
         30 . The method of  claim 24 , wherein the fetal cells are separated from the non-fetal cells by affinity chromatography. 
     
     
         31 . The method of  claim 30 , wherein the affinity chromatography is performed by contacting the cells with the one or a plurality of the antibodies that are conjugated or bound to an insoluble support. 
     
     
         32 . The method of  claim 24 , wherein the fetal cells are separated from the non-fetal cells by magnetic separation. 
     
     
         33 . The method of  claim 32 , wherein magnetic separation is performed by contacting the cells with the one or a plurality of the antibodies that are conjugated or bound to a magnetic micro-particulate support. 
     
     
         34 . The method of  claim 24 , wherein the biological fluid is maternal peripheral blood. 
     
     
         35 . The method of  claim 24 , wherein the isolated CD34+ cells are stimulated to express CD36 in the presence of:
 (i) EPO (erythropoietin), SCF, Interleukin-3 (IL-3), and Interleukin-6 (IL-6);   (ii) EPO, IGF-1 (insulin-like growth factor-1), and SCF; or   (iii) EPO, SCF, IL-3, and IGF-1.   
     
     
         36 . A method for detecting fetal cells in a biological fluid comprising the steps of:
 (a) isolating CD34+ cells from the biological fluid;   (b) stimulating the isolated CD34+ cells to express CD36;   (c) binding or labeling the cells with one or a plurality of antibodies that specifically bind to a cellular antigen that is detectably expressed by CD36+ fetal cells, but not detectably expressed by CD36+ maternal peripheral blood cells; and   (d) detecting the labeled fetal cells.   
     
     
         37 . The method according to  claim 36 , wherein the biological fluid is maternal peripheral blood. 
     
     
         38 . The method of  claim 36 , wherein the CD34+ cells are incubated in the presence of the cytokines Flt-3 ligand, TPO (thrombopoietin), and SCF (stem cell factor). 
     
     
         39 . The method of  claim 36 , wherein the isolated CD34+ cells are stimulated to express CD36 in the presence of:
 (i) EPO (erythropoietin), SCF, Interleukin-3 (IL-3), and Interleukin-6 (IL-6);   (ii) EPO, IGF-1 (insulin-like growth factor-1), and SCF; or   (iii) EPO, SCF, IL-3, and IGF-1.   
     
     
         40 . The method of  claim 36 , wherein the fetal cells are detected by flow cytometry, microscopy or radiography. 
     
     
         41 . A method for detecting fetal cells in a biological fluid comprising:
 (a) isolating CD34+ cells from the biological fluid;   (b) labeling the isolated CD34+ cells by contacting the cells with one or a plurality of detectably-labeled probes that detect expression of one or a plurality of genes that are AD026, AD037, AKNA, ASGR2, C1QG, CREM, CSPG2, CXCL16, DCNP1, GPR84, HRB2, JDP2, KCN J2, MAFB, MGC21854, MRC1, MS4A4A, MS4A6A, MS4A7, MS4A10, MS4A5, NMES1, PAG, PARVG, PKIB, PRAM-1, PTGFRN, RASGRP4, RCP, S100A8, S100A9, or TIM3; and   (c) detecting the labeled cells,   
       wherein the detected labeled cells are fetal cells. 
     
     
         42 . The method of  claim 41 , wherein the detectably-labeled probe is an antibody that specifically binds to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9 or SEQ ID NO:10. 
     
     
         43 . The method of  claim 42 , wherein the detectably-labeled probe is an antibody that specifically binds to SEQ ID NO:1. 
     
     
         44 . The method of  claim 41 , wherein the detectably-labeled probe is an mRNA probe or a riboprobe. 
     
     
         45 . The method of  claim 41 , wherein the biological fluid is maternal peripheral blood. 
     
     
         46 . The method of  claim 41 , wherein the CD34+ cells are incubated in the presence of the cytokines Flt-3 ligand, TPO (thrombopoietin), and SCF (stem cell factor). 
     
     
         47 . The method of  claim 41  further comprising, following step (a), stimulating the isolated CD34+ cells to express CD36. 
     
     
         48 . The method of  claim 47 , wherein the isolated CD34+ cells are stimulated to express CD36 in the presence of:
 (i) EPO (erythropoietin), SCF, Interleukin-3 (IL-3), and Interleukin-6 (IL-6);   (ii) EPO, IGF-1 (insulin-like growth factor-1), and SCF; or   (iii) EPO, SCF, IL-3, and IGF-1.   
     
     
         49 . The method of  claim 41 , wherein the fetal cells are detected by flow cytometry, microscopy or radiography. 
     
     
         50 . A diagnostic method wherein fetal cells obtained in accordance with  claim 24  are genetically evaluated using FISH, PCR or real time PCR.