University of Minnesota

The United States of America As Represented by the Department of Health

Biomira USA Inc.

The United States of America As Represented by the Secretary of

The United States of America As Represented by the Secretary, Department

Biormia USA Inc.

Augusto C Ochoa

Dan L Longo

Cynthia M Loeffler

Howard A Young

Paritosh Ghosh

Peter M Anderson

Arnold S Leonard

John J O'Shea

Fritz H Bach

Mircea C Popescu

Paul M Sondel

Richard J Robb

Walter Urba

Robin Lee Geller

Larry Kwak

Mark L Saxton

Mary Neville

Hiromoto Mizoguchi

Larry Boni

Barbara Alter

Hiromoto Mizuguchi

A vaccine comprising a liposome preparation including at least one B-cell malignancy-associated antigen, IL-2, alone or in combination with at least one other cytokine, and at least one type of lipid molecule, is useful in a method of inducing humoral and cellular immune responses against malignant B-cells in a mammal.

Vaccine for B-cell malignancies

Methods of identifying a patient having an altered immune status involve determining an immune status index for the patient and comparing it to the immune status index in healthy individuals. In general, an immune status index is the ratio of the amount of a protein that varies significantly in a patient with an altered immune status to the amount of another protein that is substantially invariant in both healthy and immune-altered individuals. Variable proteins can be TCR subunit proteins, T lymphocyte signal transduction pathway proteins, polynucleotide binding proteins or biological response modifiers (BRM). In addition, the ratio of a TH-1-type BRM to a TH-2-type BRM, the ratio of cytoplasmic to nuclear levels of polynucleotide binding proteins, the pattern of protein binding to an oligonucleotide probe that comprises the protein binding region of a gene for a BRM, or the pattern of distribution of T lymphocytes in a density gradient following density gradient centrifugation are also suitable as an immune status index. The methods are useful in identifying patients exhibiting immunosuppression, hyperimmunity and autoimmunity, as well as in assessing the immune status of a patient undergoing organ transplant.

Methods of identifying patients having an altered immune status

A soluble immunosuppressive factor present in serum derived from tumor-bearing mammals, is associated with changes in TCR protein subunit levels, T lymphocyte signal transduction pathway proteins. These changes provide a method of determining the level of immunosuppression in a mammal by determining the level of expression of at least one selected TCR subunit protein, a protein in the T lymphocyte signal transduction pathway, or of the NF-.kappa.B/rel family and comparing the level and pattern to that found in non-immunosuppressed individuals. The method is useful to identify patients having T lymphocytes capable of activation for immunotherapy and for identifying agents which cause or reverse immunosuppression. An isolated immunosuppressive factor associated with the level of expression of the proteins is useful for suppressing the immune response, for example, in organ transplantation.

Evaluation and treatment of patients with progressive immunosuppression

The present invention provides a liposome comprising an effective immunoadjuvant amount of a lymphokine such as IL-2. Also provided is an effective antineoplastic amount of IL-2 liposomes in combination with adoptively transferred cells stimulated with anti-CD3 monoclonal antibody plus IL-2.

Lipsome containing IL-2

The present invention provides a method for enhancing the immunotherapeutic activity, e.g., antitumor activity, of immune cells by depleting immune cells of a cell subset that down-regulates the immune response, such as either CD4.sup.+ or CD8.sup.+ lymphocytes. The remaining depleted immune cell population or the separated immune cell subsets then are cultured in the presence of an antibody to a lymphocyte surface receptor, preferably an anti-CD3 monoclonal antibody (MoAb), optionally in the presence of a relatively minor amount of interleukin-2 (IL-2). These stimulated cells then are optionally additionally cultured in the presence of IL-2 without an antibody to a lymphocyte surface receptor. The present invention also provides a method of treating a mammal having tumors or immunizing a mammal against tumors by administering the stimulated depleted immune cell population or a stimulated immune cell subset to a mammal, advantageously together with an immunosuppressant, and with liposomal IL-2. The invention further provides a method of transferring the immunity of mammals that are treated or immunized in accordance with the invention by extracting splenocytes from these mammals and administering these splenocytes to a second mammal.

Method of treating tumors with CD8.sup.+ -depleted or CD4.sup.+ T cell

Liposome immunoadjuvants containing IL-2

The present invention provides a method for enhancing the immunotherapeutic activity, e.g., cytotoxicity, of immune cells by depleting immune cells of a cell subset that down-regulates the immune response, such as either CD4.sup.+ or CD8.sup.+ lymphocytes. The remaining depleted immune cells are then cultured in the presence of interleukin-2 (IL-2) and an antibody to a lymphocyte surface receptor, preferably an anti-CD3 monoclonal antibody (MoAb). The present invention also provides a method of enhancing the immunotherapeutic activity, e.g., cytotoxicity, of immune cells by culturing immune cells in the presence of IL-2 and an antibody to a lymphocyte surface receptor, preferably an anti-CD3 MoAb; separating a cell subset capable of developing immunotherapeutic activity, e.g., cytotoxicity, from the cultured immune cells, e.g., either CD4.sup.+ or CD8.sup.+ lymphocytes; and then subculturing the separated lymphocytes in the presence of IL-2.

Method of enhancing the immunotherapeutic activity of immune cells by

A soluble immunosuppressive factor present in serum derived from tumor-bearing mammals, is associated with changes in TCR protein subunit levels and T-lymphocyte signal transduction pathway proteins. These changes provide a method of determining the level of immunosuppression in a mammal by determining the level of expression of at least one selected TCR subunit protein, or a protein in the T lymphocyte signal transduction pathway, and comparing the level to that found in non-immunosuppressed individuals. The method is useful to identify patients having T lymphocytes capable of activation for immunotherapy and for identifying agents which cause or reverse immunosuppression. An isolated immunosuppressive factor associated with the level of expression of the proteins is useful for suppressing the immune response, for example, in organ transplantation.

In accordance with the present invention, a method of culturing lymphocytes is disclosed in which the lymphocytes are cultured in the presence of interleukin-2 (IL-2) and an antibody to a lymphocyte surface receptor, preferably an antibody to a lymphocyte T3 surface receptor. Preferably, the antibody is monoclonal. Cells cultured with anti-CD3 maintain their LAK activity as they increase in number. The cells can also be cultured in the presence of an additional lymphokine to obtain additional LAK activity. Methods of medical treatment are also disclosed in which cells cultured in accordance with the culturing methods of the present invention are introduced into the individual to be treated.

Method of culturing lymphocytes and method of treatment using such

The present invention is directed to a method of increasing the in vivo immune response of lymphocytes by stimulating the lymphocytes in vitro in the presence of an anti-CD3 monoclonal antibody for less than about 24 hours to form stimulated lymphocytes; infusing the stimulated lymphocytes into a tumor-bearing mammal; and administering IL-2 to the mammal. As a result of this method, the anti-CD3 stimulated lymphocytes display enhanced immunotherapeutic, e.g., cytotoxicity or lymphokine production, in vivo as represented by a decrease in the number of tumors by at least about 20%.

Short-term anti-CD3 stimulation of lymphocytes to increase their in vivo

A method of determining the level of immunosuppression in a mammal involves determining the level of expression of at least one selected TCR subunit protein, or protein in the T lymphocyte signal transduction pathway, and comparing the level to that found in healthy individuals. The method is useful to identify patients having T lymphocytes capable of activation for autologous adoptive immunotherapy and for identifying agents which cause or reverse immunosuppression.

Growing community of inventors

Frederick, MD, United States of America

Augusto C Ochoa