Antigen‐Specific apoptosis in immortalized T cells by soluble MHC class II‐peptide complexes

Arimilli, Subhashini; Mumm, John B.; Nag, Bishwajit

doi:10.1038/icb.1996.13

Cited by 21 publications

(4 citation statements)

References 40 publications

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“…The transformed SS8T cells utilized in this study have been characterized for their restriction to HLA-DR2 and MBP(84-102) peptide (22). In our laboratory, we have shown that SS8T cells can be equally recognized by DR2 and MBP(83-102)Y 83 peptide (23,29). Results presented in showed no signi cant increase in c -IFN production (Fig.…”

Section: Resultsmentioning

confidence: 81%

Peptide Binding Inhibits Aggregation of Soluble MHC Class II in Solution

et al. 1999

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SummaryAf nity-puri ed major histocompatability complex (MHC) class II molecules are known to bind antigenic peptide in vitro. This peptide-bound MHC class II is known to undergo a change in structure upon stable binding of antigenic peptide. Previous results from our, and other laboratories, have suggested a relationship between MHC class II structure and peptide association that enables class II to enter into a stable conformation upon peptide binding. INTRODUCTIONMajor histocompatibility complex (MHC) 2 class II antigens are heterodimeric transmembrane cell surface glycoproteins that bind antigenic peptides and display them on the cell surface (1, 2). These complexes of MHC class II with antigenic peptide are then speci cally recognized by T cell receptors on the surface of T cells, leading to the MHC-restricted T cell activation (3, 4).MHC class II molecules are heterodimers with a molecular mass of 60 kDa, consisting of a 32-kDa a chain and 28-kDa b chain. The a and b chains are structurally homologous to one another, both being type I integral membrane proteins with small cytoplasmic domain and large extracellular domains. It has been documented that MHC class II proteins isolated from B lymphoblastoid cells is occupied with a wide spectrum of peptides (5-7). Several studies from different laboratories have shown that puri ed MHC class II molecules isolated from lymphoblastoid cells are capable of binding antigenic peptide in vitro (8-11) or in vivo (5-7, 12). In vitro, the percent occupancy of MHC class II molecules with antigenic peptides varies significantly. However, previous results from our laboratory showed that in vitro af nity-puri ed HLA-DR2 molecules could be fully loaded with high-af nity antigenic peptide under optimized conditions (13).Several studies with MHC class II molecules have been performed in vitro to understand a relationship between MHC class II structure and peptide binding. For example, class II molecules solubilized from cell surfaces with detergent show improved stability upon peptide binding in sodium dodecyl sulfate (SDS) (14). Similarly, baculovirus-expressed soluble human DR1 class

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Section: Resultsmentioning

confidence: 81%

Peptide Binding Inhibits Aggregation of Soluble MHC Class II in Solution

et al. 1999

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Section: Resultsmentioning

confidence: 84%

Peptide Binding Inhibits Aggregation of Soluble MHC Class II in Solution

Arimilli¹,

Astafieva²,

Mukku³

et al. 1999

IUBMB: Life

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Affinity-purified major histocompatability complex (MHC) class II molecules are known to bind antigenic peptide in vitro. This peptide-bound MHC class II is known to undergo a change in structure upon stable binding of antigenic peptide. Previous results from our, and other laboratories, have suggested a relationship between MHC class II structure and peptide association that enables class II to enter into a stable conformation upon peptide binding. In this report we describe that stable binding of high-affinity antigenic peptide to MHC class II molecule results in transition of aggregated purified MHC class II proteins to a stable heterodimeric state. Such transition was demonstrated by using purified human HLA-DR2 class II molecule and high-affinity myelin basic protein (MBP) 83-102)Y83 peptide. Highly aggregated purified DR2 (high molecular weight; HMW) was first separated from heterodimer (low molecular weight: LMW) in the presence of 50-fold molar excess of MBP(83-102)Y83 peptide. We then show that the aggregated HMW preparation can be successfully converted into a stable dimer by further incubation with MBP(83-102)Y83 and changing various binding parameters such as pH, temperature, reducing agent, and peptide concentrations. Under optimized conditions, the highly aggregated inactive DR2 molecules can be completely loaded with the antigenic peptide. The transformed heterodimers with bound peptide prepared by this method are biologically active, as shown by their ability to induce the production of gamma-interferon by SS8T-transformed human T cells. These results suggest that in solution, MHC class II molecules may be aggregated in the absence of bound peptide. Such aggregated MHC class II molecules can be converted to stable and biologically active heterodimers in the presence of high-affinity antigenic peptide.

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“…In addition, a primary hepatocyte system using enzymatic activity as an endpoint has several problems such as obtaining a stable cell preparation and reproducibility of data. In this study, we designed an in vitro assay for Kupffer cell-mediated cytotoxicity, applying the BrdU-labeled cells used for cytotoxicity assays (Ito et al, 1996;Arimilli et al, 1996). Our assay showed a good correlation between the number of co-cultured Kupffer cells and cytotoxicity.…”

Section: Discussionmentioning

confidence: 99%

Kupffer Cell- Mediated Cytotoxicity Induced by Lipopolysaccharide0111:b4 Is Greater in Dogs Than in Rats and Monkeys

Hamano

Tong²,

Mutai³

et al. 2002

J. Toxicol. Sci.

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-To clarify the mechanism of sensitivity to an endotoxin lipopolysaccharide LPS0111:B4, which causes severe liver injury in a variety of animals, we have developed an in vitro assay to measure Kupffer cell-mediated cytotoxicity in the human liver cell line, WRL68. This assay could detect the decrease in Kupffer cell activity induced by gadolinium chloride (GdCl 3 ), which is an inhibitor in Kupffer cells. Among Kupffer cells derived from dogs, rats, and monkeys, LPS-activated canine Kupffer cells exhibited remarkably high cytotoxicity against WRL68 cells. This species difference is correlated with a species difference in the lethality of LPS0111:B4. Additionally, the conditioned medium of LPS-activated canine Kupffer cells was also cytotoxic to WRL68 cells. To identify the mediators of this cytotoxicity, we measured the accelerated release of interleukin-1β, and interleukin-6 from Kupffer cells on stimulation with LPS0111:B4. From the correlation of the response to LPS0111:B4, interleukin-1β and interleukin-6 are considered to be responsible for the canine Kupffer cell-mediated cytotoxicity of LPS0111:B4.

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Antigen‐Specific apoptosis in immortalized T cells by soluble MHC class II‐peptide complexes

Cited by 21 publications

References 40 publications

Peptide Binding Inhibits Aggregation of Soluble MHC Class II in Solution

Peptide Binding Inhibits Aggregation of Soluble MHC Class II in Solution

Peptide Binding Inhibits Aggregation of Soluble MHC Class II in Solution

Kupffer Cell- Mediated Cytotoxicity Induced by Lipopolysaccharide0111:b4 Is Greater in Dogs Than in Rats and Monkeys

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