Mouse monoclonal antibodies raised to the T-cell line HSB-2 and

Details Mouse monoclonal antibodies raised to the T-cell line HSB-2 and Mouse monoclonal antibodies raised to the T-cell line HSB-2 and

1990-05-23

1992-10-27

Doll, John J.

Chemistry: molecular biology and microbiology

Spore forming or isolating process

5303887, 53038873, 53038875, C12N 520, C07K 1528

Patent

active

051588854

DESCRIPTION:

BRIEF SUMMARY
FIELD OF THE INVENTION

The present invention relates to two new hybridoma cell lines and to monoclonal antibodies produced by these hybridoma cell lines. Both these monoclonal antibodies exhibit broad range reactivity with human leucocytes, and have potential for use as immunosuppressive agents.


BACKGROUND OF THE INVENTION

Since it was shown by Kohler and Milstein (Nature Vol. 256, 495-497, 1975) that it was possible to fuse mouse myeloma cells with spleen cells from immunized mice and thereby produce a continuous cell line which produces a homogeneous (monoclonal) antibody, extensive attention has been focused on the production of these hybrid cell lines (hybridomas) and the monoclonal antibodies (Mabs) produced.
The development of hybridoma technology has led to a drammatically improved understanding of the antigenic molecules on the surface of human leucocytes. Much of this information has recently been systematised in the International Workshop on Human Leucocyte Differentiation Antigens, and the majority of many thousands of Mabs reported can now be recognised as belonging to one of the 45 accepted Clusters of Differentiation (CD). The majority of these clusters define antigens which are restricted to specific leucocyte differentiation lineages or to maturation stages. One group of Mabs, however, belonging to CD 45, define a 200 kilodalton protein, now known as Leucocyte Common Antigen (LCA) or T-200, which has the unusual property of being expressed on virtually on human leucocytes.
There are only a few other molecules with a similar distribution. Class I histocompatibility antigens are found on all leucocytes (with the exception of immature thymocytes), but are also expressed on a wide variety of non-haemopoietic tissue. Two other antigens, identified by Mabs CAMPATH-1 (Blood, 1983; 62:873) and HuLyM3 (Transplantation; 1983 36:446) also have pan-haemopoietic distribution, although their expression on non-haemopoietic cells is less certain.


SUMMARY OF THE INVENTION

In a first aspect the present invention consists in a mouse monoclonal antibody of Class IgM produced by a hybridoma formed by fusion of cells from a mouse myeloma line and spleen cells from a mouse previously immunized with the T cell line HSB-2 and T-CLL cells, the monoclonal antibody being characterised in that it reacts with leucocyte differentiation antigen of a relative molecular weight of approximately 65 kilodaltons which is expressed on over 90% of normal human peripheral blood mononuclear cells but not on normal granulocytes, platelets or erythrocytes, and that it is essentially unreactive with human leukaemic cells either from fresh cases of acute leukaemia or the commonly available leukaemic cell lines of B, T and myeloid derivation.
It is preferred that the monoclonal antibody is further characterised in that it:
(a) Reacts with approximately 95% of normal human thymocytes;
(b) Reacts with greater than 97.5% of tonsil lymphocytes;
(c) Reacts with approximately 95% of activated human lymphocytes;
(d) Fixes both human and rabbit complement; and
In a second aspect the present invention consists in an IgM monoclonal-antibody-producing hybridoma cell line formed by fusion of cells from a mouse myeloma line and spleen cells from a mouse previously immunized with the T cell line HSB-2 and T-CLL cells, the monoclonal antibody produced being characterised in that it reacts with a leucocyte differentiation antigen of a relative molecular weight of approximately 65 kilodaltons which is expressed on over 90% of normal human peripheral blood mononuclear cells but not on normal granulocytes, platelets or erythrocytes and that it is essentially unreactive with human leukaemic cells either from fresh cases of acute leukaemia or the commonly available leukaemic cell lines of B,T and myeloid derivation.
It is preferred that the monoclonal antibody produced is further characterized in that it:
(a) Reacts with approximately 95% of normal human thymocytes;
(b) Reacts with greater than 97.5% of tonsil lymphocytes;
(c) Reacts with approximate

REFERENCES:
Becker et al., "Monoclonal Antibodies to Human Macrophage and Leucocyte Common Antigens," Pathology, 13, 669-680 (1981).
Vaughan et al., "Hu Ly-m3-A Human Leukocyte Antigen," Transplantation, 36, 446-450 (1983).
Hale et al., "Removal of T Cells from Bone Marrow for Transplantation: A Monoclonal Antilymphocyte Antibody That Fixes Human Complement," Blood, 62, 873-882 (1983).

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