Composition for stabilizing survival of transplanted hematopoietic stem cell, kit for obtaining the composition, method of stabilizing survival of transplanted hematopoietic stem cell, human monoclonal antibody or human polyclonal antibody and method of producing the same, gene encoding human monoclonal antibody and transf

Abstract

[PROBLEMS] To stabilize the survival of transplanted hematopoietic stem cells by using HLA identical activated lymphocytes. [MEANS FOR RESOLVING PROBLEMS] HLA identical activated lymphocytes in mononuclear cells separated from peripheral blood or umbilical cord blood are proliferated and activated. After separating and collecting, the HLA identical activated lymphocytes are employed as the main component of a composition for promoting survival of transplanted hematopoietic stem cells. The obtained composition is widely usable in, for instance, prevention of survival failure of transplanted hematopoietic stem cells and therapy for promoting the survival thereof. Although the dose of the composition varies depending on the age, conditions, etc. of a patient, a humanized antibody is administered in a dose of from 0.2 to 20 ml/kg/day to mammals including humans. The composition is administered by intravenous injection either once a day (single administration or continuous administration) or intermittently once to 3 times in a week or once in 2 or 3 weeks.

1 . A composition for promoting survival after hematopoietic stem-cell transplantation, wherein: after HLA identical lymphocytes in mononuclear cells separated from peripheral blood or umbilical cord blood are proliferated and activated, said HLA identical lymphocytes separated and collected from the resulting matter are employed as a main component.  
   
   
       2 . A composition for promoting survival after hematopoietic stem cell transplantation according to  claim 1 , wherein are contained said HLA identical lymphocytes as a main component and a component for treatment.  
   
   
       3 . A composition for promoting survival after hematopoietic stem-cell transplantation according to  claim 1 , wherein said HLA identical activated lymphocytes are CD4+ cells.  
   
   
       4 . A kit for obtaining a composition for promoting survival after hematopoietic stem cell transplantation, wherein said kit is an instrument for proliferating and activating HLA identical lymphocytes collected from peripheral blood or umbilical cord blood, and said instrument comprises a culture medium containing interleukin 2 and an anti-CD3 antibody solidified flask.  
   
   
       5 . A method for producing a composition for promoting survival after hematopoietic stem cell transplantation, comprising a process for separating mononuclear cells from peripheral blood or umbilical cord blood containing HLA identical lymphocytes; a process for proliferating and activating HLA identical lymphocytes in said separated mononuclear cells; and a process for separating said proliferated and activating HLA identical lymphocytes and preparing them to a formulation in which they constitute a main component.  
   
   
       6 . A method for producing a composition for promoting survival after hematopoietic stem cell transplantation according to  claim 5 , wherein proliferation and activation of said HLA identical lymphocytes are carried out in any one of interleukin 2 or anti-CD3 antibody or in combination of them.  
   
   
       7 . A method for producing a composition for promoting survival after hematopoietic stem-cell transplantation according to  claim 5  or  6 , wherein various kinds of cytokines or mitogens are added to medium for proliferation and activation of said HLA identical lymphocytes.  
   
   
       8 . A human monoclonal antibody producing cell lines collected by a method for administering HLA identical activated lymphocytes before or after transplanting to an immunodeficiency animal, immunizing said animal to antigens after promoting survival, establishing or separating antigen sensitized lymphocytes from said animal, or gene separation or manifestation.  
   
   
       9 . A human polyclonal antibody produced by transplanting human hematopoietic stem cells to an immunodeficiency animal by administering HLA identical lymphocytes to said animal before or after the transplantation, by immunizing antigens after promoting survival, and collected from said animal.  
   
   
       10 . A method for establishing human monoclonal antibody producing cell lines, wherein said human monoclonal antibody producing cell lines are collected by a method that human hematopoietic stem cells are transplanted to an immunodeficiency animal, survival of them is promoted by administering HLA identical lymphocytes before or after the transplantation and immunizing animal with the antigen, said sensitized lymphocytes are established as a line or separated, or collected by a method of gene separation or gene expression manifestation.  
   
   
       11 . A method for producing human monoclonal antibody producing cell lines according to  claim 10 , wherein said immunodeficiency animal is a severe combined immunodeficiency mouse.  
   
   
       12 . A method for producing human monoclonal antibody producing cell lines according to  claim 11 , wherein said severe combined immunodeficiency mouse is a SCID mouse.  
   
   
       13 . A method for producing human monoclonal antibody producing cell lines according to  claim 10 , wherein said administered HLA identical human lymphocytes are HLA identical human activated lymphocytes.  
   
   
       14 . A method for producing human monoclonal antibody producing cell lines according to  claim 10 , wherein said administered HLA identical human activated lymphocytes contain CD4+ cells as a main component.  
   
   
       15 . A method for producing human monoclonal antibody producing cell lines according to  claim 10 , wherein said administered HLA identical human activated lymphocytes contain not less than 90% of activated CD4+ cells relative to total administered cell number.  
   
   
       16 . A method for producing human monoclonal antibody producing cell lines according to  claim 10 , wherein contamination rate of CD8+ cells relative to said administered HLA identical human activated lymphocytes is not more than 10% relative to total administered cell number.  
   
   
       17 . A method for producing human monoclonal antibody producing cell lines according to  claim 10 , wherein a means for establishing said antigen sensitized lymphocytes from the immunodeficiency animal is a way of establishing with Epstein-Barr virus.  
   
   
       18 . A method for producing human monoclonal antibody producing cell lines according to  claim 10 , wherein a means for establishing said antigen sensitized lymphocytes from the immunodeficiency animal is a way of establishing by cell fusion with human myeroma cell line.  
   
   
       19 . A method for producing human monoclonal antibody producing cell lines according to any one of  claim 10  to  18 , wherein said antigen is a human-derived substance.  
   
   
       20 . A method for producing human monoclonal antibody producing cell lines according to any one of  claim 10  to  18 , wherein said antigen is a complex of human-derived hapten and carrier protein.  
   
   
       21 . A method for producing human monoclonal antibody producing cell lines according to any one of  claim 10  to  18 , wherein said antigen is MAP (Multiple Antigen Peptide) composed of a human-derived peptide.  
   
   
       22 . A method for producing human monoclonal antibody producing cell lines according to any one of  claim 10  to  18 , wherein said antigen is an immunogloblin.  
   
   
       23 . A method for producing human monoclonal antibody producing cell lines according to  claim 10 , wherein said antigen is an IgG immunogloblin.  
   
   
       24 . A human monoclonal antibody produced by human monoclonal antibody producing cell lines according to any one of  claim 8 ,  10  to  23 .  
   
   
       25 . A gene coding a human monoclonal antibody produced by human monoclonal antibody producing cell lines according to any one of  claim 8 ,  10  to  23 .  
   
   
       26 . A tranformant introduced a gene coding a human monoclonal antibody produced by human monoclonal antibody producing cell lines according to any one of  claim 8 ,  10  to  23 .  
   
   
       27 . A method for producing human polyclonal antibody produced by transplanting human hematopoietic stem cells to an immunodeficiency animal, promoting survival by administering HLA identical lymphocytes before or after the transplantation and immunizing with antigen, and collecting from said immunodeficiency animal.  
   
   
       28 . A method for producing human polyclonal antibody according to  claim 27 , wherein said administered HLA identical lymphocytes are HLA identical activated lymphocytes.  
   
   
       29 . A method for producing human polyclonal antibody according to  claim 27 , wherein said administered HLA identical human activated lymphocytes contain CD4+ cells as a main component.  
   
   
       30 . A method for producing human polyclonal antibody according to  claim 27 , wherein said administered HLA identical human activated lymphocytes contain not less than 90% of activated CD4+ cells relative to total administered cell number.  
   
   
       31 . A method for producing human polyclonal antibody according to  claim 27 , wherein contamination rate of CD8+ cells relative to said administered HLA identical human activated lymphocytes is not more than 10% relative to total administered cell number.  
   
   
       32 . A method for producing human polyclonal antibody producing cell line according to any one of  claim 27  to  31 , wherein said antigen is a human-derived substance.  
   
   
       33 . A method for producing human polyclonal antibody producing cell line according to any one of  claim 27  to  31 , wherein said antigen is a complex of human-derived hapten and carrier protein.  
   
   
       34 . A method for producing human polyclonal antibody producing cell lines according to any one of  claim 27  to  31 , wherein said antigen is MAP (Multiple Antigen Peptide) composed of a human-derived peptide.  
   
   
       35 . A method for producing human polyclonal antibody producing cell lines according to any one of  claim 27  to  31 , wherein said antigen is an immunogloblin.  
   
   
       36 . A method for producing human monoclonal antibody producing cell lines according to any one of  claim 27  to  31 , wherein said antigen is an IgG immunogloblin.  
   
   
       37 . A human polyclonal antibody produced by human monoclonal antibody producing cell lines according to any one of  claim 9 ,  27  to  36 .