Lymphocyte production method

Inventors

Otsuji, Tomomi • HIRASE, Yuka • Hatsuyama, Asako • Okamoto, Sachiko • Enoki, Tatsuji • Mineno, Junichi

Assignees

Takara Bio Inc

Abstract

In the present invention, lymphocytes are efficiently grown by culturing lymphocytes in the presence of a novel recombinant fibronectin fragment.

Core Innovation

The invention relates to a method of producing a lymphocyte by culturing a lymphocyte in the presence of recombinant fibronectin fragments comprising human fibronectin type III repeats. The method uses recombinant polypeptides corresponding to fibronectin III-1 to III-3, fibronectin III-8 to III-10, and fibronectin III-12 to III-14, wherein the recombinant polypeptides are not full-length human fibronectin.

The background problem addressed is that prior fibronectin-fragment approaches mainly yield cytotoxic T cells rather than maintaining or enriching helper T cells, and therefore do not provide lymphocyte products with the desired helper/naive-like characteristics. The disclosed method is described as efficiently expanding lymphocytes with maintained function, with an emphasis on CD4-expressing helper T cells.

In the disclosed approach, culturing in the presence of the specified recombinant fibronectin type III repeats increases the proportion of CD4-expressing cells compared with culturing in the absence of these recombinant polypeptides. The method is also described as enabling helper T cells having a naive-like phenotype, including CD45RA+ CCR7+ cells, and can be performed with additional culture components such as anti-CD3 and solid-phase coated culture devices.

Claims Coverage

The independent claim coverage centers on a lymphocyte production method that uses three specific sets of recombinant human fibronectin type III repeats (1-3, 8-10, and 12-14) as non-full-length fragments, and requires the resulting lymphocytes to have a higher proportion of CD4-expressing cells than cultures without these fragments. The claim set includes refinements that combine the repeat-containing polypeptides into one molecule, specify SEQ ID NO: 19, add anti-CD3, and select among solid-phase coated culture device options.

Overall, the claim coverage focuses on culturing lymphocytes with non-full-length recombinant human fibronectin type III repeat fragments (repeats 1-3, 8-10, and 12-14) to obtain lymphocytes with a higher proportion of CD4-expressing cells, with refinements that include combining segments into a single construct, specifying SEQ ID NO: 19, adding anti-CD3, and selecting from listed solid-phase forms.

Stated Advantages

Documented Applications