25 attenuation of lesion tissue; and 5) preservation of nerve cells and axons in presence of a highly variable astroglial reaction. These changes are not in support of a pathogenesis of primary CNS tissue damage by HIV. Rather, they suggest secondary pathogenetic events involving humoral factors (vasogenic edema following focal blood barrier disruption, or local action of mediators of inflam- mation or immunoglobulins) and/or cellular effectors (cell- mediated immune attack on myelin triggered by HIV, or bystander demyelination by locally present macrophages). An immunopatho- logic or bystander damage of myelin triggered by HIV might be comparable to inflammatory demyelination found in brains infected with the closely related visna-maedi retrovirus. Isolation of Human Lymphocyte Cell Lines Reactive with Whole Human Myelin James Burns and Kimberly Littlefield Department of Neurology University of Utah School of Medicine 50 North Medical Drive Salt Lake City, Utah 84132 Multiple sclerosis (MS) may be the result of an autoimmune response directed against CNS myelin. A possible animal model for MS, experimental allergic encephalomyelitis (EAE), may be induced by immunization with purified components of whole myelin including myelin basic protein (MBP), proteolipid protein, and DM-20. We and others have isolated CD4+ T-cells reactive with MBP from the peripheral blood lymphocytes (PBL) of human subjects by in vitro sensitization with purified MBP. We now have used whole human myelin and delipidated myelin for in vitro sensitization to attempt the identification of antigens other than MBP which may be recognized by human T- lymphocytes. Long term cell lines reactive with myelin or delipidated myelin were established from the PBL of a normal subject. These cell lines were maintained in culture for two months by repeated stimulation with the crude myelin antigens. Eventhough these cells were not exposed to purified MBP, the cell lines proliferated briskly when cultured with MBP. Cells reactive with delipidated myelin were cloned with six of eight clones responding to MBP as well as delipidated myelin and two clones poorly reactive with either antigen. For this normal subject, under the culture conditions employed, MBP appeared to be the major antigen recognized in delipidated myelin. These results suggest that whole myelin may be used for in vitro sensitization to study the human cellular immune response to myelin antigens.

Isolation of human lymphocyte cell lines reactive with whole human myelin

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Page 1: Isolation of human lymphocyte cell lines reactive with whole human myelin

attenuation of lesion tissue; and 5) preservation of nerve cells

and axons in presence of a highly variable astroglial reaction.

These changes are not in support of a pathogenesis of primary CNS

tissue damage by HIV. Rather, they suggest secondary pathogenetic

events involving humoral factors (vasogenic edema following focal

blood barrier disruption, or local action of mediators of inflam-

mation or immunoglobulins) and/or cellular effectors (cell-

mediated immune attack on myelin triggered by HIV, or bystander

demyelination by locally present macrophages). An immunopatho-

logic or bystander damage of myelin triggered by HIV might be

comparable to inflammatory demyelination found in brains infected

with the closely related visna-maedi retrovirus.

Isolation of Human Lymphocyte Cell Lines Reactive with Whole Human Myelin

James Burns and Kimberly Littlefield Department of Neurology University of Utah School of Medicine 50 North Medical Drive Salt Lake City, Utah 84132

Multiple sclerosis (MS) may be the result of an autoimmune response directed against CNS myelin. A possible animal model for MS, experimental allergic encephalomyelitis (EAE), may be induced by immunization with purified components of whole myelin including myelin basic protein (MBP), proteolipid protein, and DM-20. We and others have isolated CD4+ T-cells reactive with MBP from the peripheral blood lymphocytes (PBL) of human subjects by in vitro sensitization with purified MBP. We now have used whole human myelin and delipidated myelin for in vitro sensitization to attempt the identification of antigens other than MBP which may be recognized by human T- lymphocytes. Long term cell lines reactive with myelin or delipidated myelin were established from the PBL of a normal subject. These cell lines were maintained in culture for two months by repeated stimulation with the crude myelin antigens. Eventhough these cells were not exposed to purified MBP, the cell lines proliferated briskly when cultured with MBP. Cells reactive with delipidated myelin were cloned with six of eight clones responding to MBP as well as delipidated myelin and two clones poorly reactive with either antigen. For this normal subject, under the culture conditions employed, MBP appeared to be the major antigen recognized in delipidated myelin. These results suggest that whole myelin may be used for in vitro sensitization to study the human cellular immune response to myelin antigens.