Search PubMed⌕ Search

Biomedical subjects

Q J Zhang

Publications and source records attributed to Q J Zhang.

At least 19 recordsLinked to original sources
PubMed2001

Defect in insulin-like growth factor-1 survival mechanism in atherosclerotic plaque-derived vascular smooth muscle cells is mediated by reduced surface binding and signaling.

Apoptosis of vascular smooth muscle cells (VSMCs) is increased in atherosclerosis compared with normal vessels, where it may contribute to plaque rupture. We have previously found that human plaque-derived VSMCs (pVSMCs) are intrinsically sensitive to apoptosis and not responsive to the protective effects of insulin-like growth factor-1 (IGF-1). We therefore examined the mechanism underlying this defect. Human pVSMCs showed <25% (125)I-IGF-1 surface binding, <20% IGF-1 receptor (IGF-1R) expression than that of normal medial VSMCs, and <40% Akt kinase activity in response to IGF-1. pVSMCs expressed and secreted high levels of IGF-1 binding proteins (IGFBPs), and the IGF-1 analogues, long R3 and Des 1,3 IGF-1, which do not bind to IGFBPs, were able to increase pVSMC survival to normal medial VSMC levels. The long R3 survival effect was phosphatidylinositol 3-kinase-mediated, but it was not dependent on Akt activity alone. Intimal pVSMCs in vivo showed reduced IGF-1R expression compared with medial VSMCs, in particular at the shoulder regions of plaques. We conclude that human pVSMCs show an intrinsic sensitivity to apoptosis caused in part by defective expression of IGF-1R, impaired IGF-1-mediated survival signaling and increased IGFBP secretion. This impaired IGF-1 protection against apoptosis may promote VSMC loss and plaque instability in atherosclerosis.

Androstadienes↗
PubMed2001

Effects of repeated +Gz exposures on lipid peroxidation of various organs in rats.

OBJECTIVE: To investigate the change of lipid peroxidation in various organs of rats after repeated +10 Gz stress. METHOD: Twenty male Wistar rats were randomly divided into control (+1 Gz) group and +10 Gz group (n = 10 for each group ). The two groups were exposed to +1 Gz or +10 Gz on an animal centrifuge respectively. The onset rate of +Gz was 0.5 G/s; the sustained time at peak +Gz was 30 s each time, with 5 times/d, 1 min intervals, 3 d/wk, 3 wk in total. The rats were decapitated in ice bath on the next day after the last +Gz exposure. The lung, heart, liver and kidney of rats were collected for the following determinations. These organs were made into homogenates and the mitochondria of heart and kidney were isolated from their homogenates. Then malondialdehyde (MDA) content , superoxide dismutase (SOD) activity and glutathion (GSH) concentration in these homogenates and mitochondria were determined individually. RESULT: Compared with the control, MDA concentration in myocardial mitochondria increased significantly (P < 0.01) after repeated +10 Gz exposures, and the SOD activity in liver homogenate and renal mitochondria decreased remarkably (P < 0. 01). But there was no significant difference between GSH concentrations of the two groups. CONCLUSION: Repeated +10 Gz stress induced lipid peroxidation injury of myocardial mitochondria and had some influences on the oxygen radical metabolism of liver and kidney in rats.

Animals↗
PubMed2001

Protective effects of tea polyphenols on myocardial free radical metabolic disorder in mice induced by inhalation of pure oxygen under 5500 m hypobaric condition.

OBJECTIVE: To observe the protective effects of natural antioxidant tea polyphenols(TP) on myocardial free radical metabolic disorder in mice induced by inhalation of hypobaric pure oxygen under 5500 m hypobaric condition. METHOD: Forty-two male Kunming mice were randomly divided into three groups (n = 14 each): group A, normal control; group B, inhalation of pure oxygen (> 96 %) at simulated altitude of 5500 m in an animal altitude chamber; group C (TP protection group), same as group B but 100 mg/kg of TP was given orally before the exposure. The exposure time was 2 h/d, 3 d/wk for a total of 8 wk, and distilled water was given to groups A and B before exposure. After experiment, the mice were decapitated on the next day and the heart was quickly removed. Malondialdehyde (MDA) concentration, superoxide dismutase (SOD) activity and nitric oxide (NO) content were measured. In addition, Cu, Zn-SOD and inducible NO synthase (iNOS) enzymatic contents in myocardial tissue were qualitatively examined by immunohistochemical assaying. RESULT: Compared with the control, MDA concentration, SOD activity and Cu, Zn-SOD enzymatic content in group B were significantly increased (P < 0. 05). But in TP protection group, myocardial MDA formation was significantly decreased (P < 0. 01) and SOD activity and Cu, Zn-SOD expression restored to normal. On the contrary, myocardial NO generation and iNOS expression were significantly reduced after repeated inhalation of hypobaric oxygen at 5500 m. NO metabolism regained to normal after repeated administration of TP. CONCLUSION: Natural antioxidant TP had protective effects on myocardial free radical metabolic disorder induced by inhalation of hypobaric pure oxygen under 5500 m hypobaric condition.

Altitude↗
PubMed2001

[Inhibitory effects of tea polyphenols and vitamin C on lipid peroxidation induced by FeSO4- cysteine in isolated human plasma and carbon tetrachloride-induced liver free radical injury in mice].

OBJECTIVE: To investigate the inhibitory effects of tea polyphenols (TP) and Vitamin C (Vit C) on FeSO4-cysteine induced lipid peroxidation in isolated human plasma and carbon tetrachloride (CCl4) induced liver free radical injury in mice. METHOD: The experiment included two parts: (1) In FeSO4-cysteine induced lipid peroxidation system of isolated human plasma, malondialdehyde (MDA) content was detected after administration of different concentrations of TP (0.3 ~ 8.1 mg/L, as C ~ F group) and Vit C (3 ~ 81 mg/L, as G ~ J group) respectively; (2) Thirty-six male Kunming mice were randomly divided into four groups: control group (A), CCl4 damage group (B), TP protection group (C) and Vit C protection group (D). TP or Vit C (100 mg/kg) was given orally to group C and D respectively, while the same volume of distilled water was given to the other two groups one time every day, continued for three days. Twelve hours after the final treatment, CCl4 (230 mg/kg) was given orally to group B ~ D. Thirty six hours later, all the mice were decapitated and liver homogenate were prepared for measuring MDA content. RESULT: In FeSO4-cysteine induced lipid peroxidation in isolated human plasma, the inhibitory rate of TP (0.3 ~ 8.1 mg/L) was 30.7%, 32.0%, 46.9% and 59.7 %, and the inhibitory rate of Vit C (3 ~ 8.1 mg/L) was 8.3%, 41.4%, 47.7% and 52.7% for various dosages. In the CCl4 induced liver free radical injury system, the inhibitory rate of the same dosage of TP and Vit C was 45.2%, 42.8% respectively. CONCLUSION: TP (0.3 ~ 8.1 mg/L) and Vit C (9 ~ 81 mg/L) had inhibited lipid peroxidation induced by FeSO4-cysteine in isolated human plasma significantly, and the inhibitory effects of TP was superior to that of Vit C at the same dosages. The same dosage of TP and Vit C had remarkable inhibitory effects on the CCl4 induced liver free radical injury, but there was no significant difference between the two groups.

Animals↗
PubMed2001

Protective effects of tea polyphenols on mild hypobaric hypoxia induced pulmonary free radical metabolic disorder in mice.

Objective. To observe the protective effects of natural antioxidant tea polyphenols (TP) on repeated mild hypobaric hypoxia induced pulmonary free radical metabolic disorder in mice. Method. Fourty-two male Kunming mice were randomly divided into three groups (n=14 each): normal control (A); 1500 in mild hypobaric hypoxia (B) and TP protection group (C). The exposure time in hypobaric chamber was 2 h/d, 3d/wk, 8 wk in total. Before hypoxic exposure, TP was orally given to group C at a dose of 100 mg/kg, while distilled water was given to the other two groups. After experiment, the mice were decapitated on the next day and the lung was quickly removed. The malondialdehyde (MDA) concentration, superoxide dismutase (SOD) activity and nitric oxide (NO) content were measured. In addition, Cu, Zn-SOD and inducible NO synthase (iNOS) enzymatic contents in lung were were qualitatively examined by immunohistochemical assaying. Result. Compared with the control group, pulmonary MDA concentration and NO content were significantly increased after chronic mild hypobaric hypoxic exposures (P<0.01) but the MDA formation and NO generation in TP protection group were restored to normal. Pulmonary SOD activity in group B tended to increase. Cu, Zn-SOD expression in endothelial cells of bronchioli and iNOS contents in endothelial cells of bronchioli and endothelial cells and smooth muscle cells in pulmonary interstitial vessels were significantly elevated after repeated mild hypobaric hypoxic exposure. These enzymatic abnormal expressions regained to normal after administration of TP. Conclusion. Natural antioxidant TP had protective effects on repeated mild hypobaric hypoxia induced pulmonary free radical metabolic disorder.

Altitude↗
PubMed2001

[Effects of repeated high +Gz exposure on several enzyme activities in cardiomyocytes in rats and some protective measures].

Objective. To determine the changes of several myocardial enzymes in rats after repeated high +Gz exposure and the protective effects of preconditioning of low-G exposure and tea polyphenols (TP). Method. Thirty-two male Wistar rats were randomly divided into 4 groups (n = 8 each): control group (group A), +10 Gz group (group B), low-G preconditioning group (group C) and TP protection group (group D). Group B, C and D were exposed to repeated +10 Gz stress (each for 30s, 5 times/d with +1 Gz 1 min intervals, 3 d/wk, 3 weeks in total), but group A was only submitted to +1 Gz for 5 min. Group C was exposed to +2 Gz for 5 min about 1 h prior to +10 Gz stress. Additionally, TP (200 mg/kg) was given orally to group D about 1 h prior to the +Gz experiment, while distilled water was given to groups A, B and C instead. On the next day after the last centrifuge run, the hearts were taken out immediately for making frozen tissue sections. Enzyme histochemical staining and image analysis were carried out for acid phosphatase (ACP), succinate dehydrogenase (SDH), cytochromeoxidase (Cyt aa3) and adenosine triphosphatase (ATPase). Result. As compared with the control, the activities of ACP and SDH in +10 Gz stressed rats decreased significantly (P<0.05), and there was a declining trend for Cyt aa3. But, low-G preconditioning and TP had protective effects on +10 Gz stress-induced reduction of these enzymatic activities (P<0.05). Conclusion. The results showed that repeated high +Gz exposure could bring about decreases of activities in ACP (the marker enzyme of lysosome) and SDH (the marker enzyme of mitochondrial endomembrane), which indicated a reduction of oxidative metabolism in myocardial tissue; but preconditioning with low-G and natural antioxidant TP had protective effects.

Acceleration↗
PubMed2001

[Responses of afferent unit of the caudal nerve of diabetic hyperalgesic rats to sympathetic efferent stimulation].

Responses of afferent unit to sympathetic stimulation (SS), intravenous injection of noradrenaline (NA) and phentolamine in the caudal nerve of diabetic rats were investigated. The results showed that the discharge frequencies of C and Adelta units with spontaneous discharges were increased in diabetic hyperalgesic rats after SS, and these spontaneous discharges were eliminated by adrenergic antagonist. The C (6/21) and Adelta (19/81) units with no spontaneous discharges of diabetic hyperalgesic rat turned from silent state into active state during SS; although SS did not elicit afferent discharges of the C mechanical receptive units (C-M), it elicited afferent discharges of a part of C mechano heat units (C-MH) and C polymodal units (C-Pol); afferent discharges of some of the Adelta mechanical receptive units (Adelta-M) and Adelta mechano heat units (Adelta-MH) were also elicited by SS. The latencies of the C and Adelta units responses upon SS were not equal, but no less than 5 s. SS elicited neither afferent discharges from Abeta mechanical receptive units of diabetic hyperalgesic rat, nor receptive units of all types in the control rat. The C(3/8) and Adelta (4/12) units of diabetic hyperalgesic rat were activated by intravenous injection of NA. The present data suggest that NA released from sympathetic nerve terminals excites C and Adelta units of diabetic hypesthesic rat, which may be a peripheral factor in hyperalgesia and paresthesia of diabetic rats.

Afferent Pathways↗
PubMed2000

Evidence of selective processing of immunodominant epitopes in virally infected cells.

Recent advances in clarifying the molecular mechanisms involved in Ag processing and presentation have relied heavily on the use of somatic cell mutants deficient in proteasome subunits, TAP transporter, and cell surface expression of MHC class I molecules. Of particular interest currently are those mutants that lack specific protease activity involved in the generation of antigenic peptides. It is theoretically possible that deficiencies of this nature could selectively prevent the cleavage of certain peptide bonds and thus generate only a subset of antigenic peptides. Gro29/Kb cell line is derived from the wild-type murine Ltk- cell line. This cell line is one example of a mutant that lacks specific protease activities. This deficiency manifests itself in an inability to generate a subset of immunodominant peptide epitopes derived from vesicular stomatitis virus and herpes simplex virus. This in turn leads to a general inability to present these viral epitopes to cytotoxic T lymphocytes (CTL). These studies describe a unique Ag processing deficiency and provide new insight into the role of proteasome-independent proteases in MHC class I-restricted peptide generation.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed2000

TAP expression provides a general method for improving the recognition of malignant cells in vivo.

A major class of tumors lack expression of the transporters associated with antigen processing (TAP). These proteins are essential for delivery of antigenic peptides into the lumen of the endoplasmic reticulum (ER) and subsequent assembly with nascent major histocompatibility complex (MHC) class I, which results in cell surface presentation of the trimeric complex to cytolytic T lymphocytes. Cytolytic T lymphocytes are major effector cells in immunosurveillance against tumors. Here we have tested the hypothesis that TAP downregulation in tumors allows immunosubversion of this effector mechanism, by establishing a model system to examine the role of TAP in vivo in restoring antigen presentation, immune recognition, and effects on malignancy of the TAP-deficient small-cell lung carcinoma, CMT.64. To test the potential of providing exogenous TAP in cancer therapies, we constructed a vaccinia virus (VV) containing the TAP1 gene and examined whether VV-TAP1 could reduce tumors in mice. The results demonstrate that TAP should be considered for inclusion in cancer therapies, as it is likely to provide a general method for increasing immune responses against tumors regardless of the antigenic complement of the tumor or the MHC haplotypes of the host.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1999

[Psychological issues in manned spaceflight].

As the duration of manned spaceflight becomes longer and as crews become more heterogeneous, psychological and interpersonal factors will be more important in affecting the safety of crew and flight mission. In space environment there are four types of stressors: physical, physiological, psychological and interpersonal. Psychological issues include "Asthenia", alteration in time sense, transcendent experiences, sleep problem, career motivation, psychosomatic symptoms and psychiatric issues. Interpersonal issues include interpersonal tension, interpersonal relationships decreased cohesiveness and deprivation, displacement [correction of dispiacement] of anger to outside personnel over time.

Adaptation, Psychological↗
PubMed1999

[Effects of simulated flight hypobaric hypoxia and oxygen inhalation on free radical metabolism in various organs of mice].

Objective. To investigate the effects of two types of simulated flight conditions (hypobaric hypoxia and hypobaric oxygen inhalation) on free radical metabolism in various organs of mice. Method. Sixty male Kunming mice were randomly divided into six groups (n=10 each). The experiment comprises two parts. The first part included three groups: normal controls (A1), 1500 m hypobaric hypoxia for 4 wk (B1) and 8 wk (C1). The second part included another three groups: normal control (A2), 5500 m hypobaric oxygen inhalation for 4 wk (B2) and 8 wk (C2). The exposure time in hypobaric chamber was 2 h/d, 3 d/wk. After experiment, caudal blood was taken for routine examination. The mice were decapitated on the next day and brain, heart, lung, liver and kidney homogenates were prepared for measuring malondialdehyde (MDA) content and superoxide dismutase (SOD) activity. Result. Lipid peroxides in lung was significantly increased in C1 group, and the content of myocardial MDA and myocardial SOD activities in C2 group were markedly higher than those in A2 group. There were no significant differences among body weights, mean corpuscular indices and hemoglobin content in the normal control, hypobaric hypoxia and hypobaric oxygen inhalation groups. It demonstrates that repeated mild hypobaric hypoxia for 8 wk causes free radical damage of lung and repeated exposure to 5500 m hypobaric oxygen inhalation for 8 wk may lead to myocardial peroxidative injury in mice. Conclusion. Simulated flight hypobaric hypoxia and oxygen inhalation may lead to free radicals damage of lung and myocardial peroxidative injury in mice.

Altitude↗
PubMed1998

Surrogate antigen processing mediated by TAP-dependent antigenic peptide secretion.

MHC class I proteins assemble with peptides in the ER. The peptides are predominantly generated from cytoplasmic proteins, probably by the action of the proteasome, a multicatalytic proteinase complex. Peptides are translocated into the ER by the transporters associated with antigen processing (TAP), and bind to the MHC class I molecules before transport to the cell surface. Here, we use a new functional assay to demonstrate that peptides derived from vesicular stomatitis virus nucleoprotein (VSV-N) antigen are actively secreted from cells. This secretion pathway is dependent on the expression of TAP transporters, but is independent of the MHC genotype of the donor cells. Furthermore, the expression and transport of MHC class I molecules is not required. This novel pathway is sensitive to the protein secretion inhibitors brefeldin A (BFA) and a temperature block at 21 degrees C, and is also inhibited by the metabolic poison, azide, and the protein synthesis inhibitor, emetine. These data support the existence of a novel form of peptide secretion that uses the TAP transporters, as opposed to the ER translocon, to gain access to the secretion pathway. Finally, we suggest that this release of peptides in the vicinity of uninfected cells, which we term surrogate antigen processing, could contribute to various immune and secretory phenomena.

ATP-Binding Cassette Transporters↗
PubMed1997

Natural variants of the immunodominant HLA A11-restricted CTL epitope of the EBV nuclear antigen-4 are nonimmunogenic due to intracellular dissociation from MHC class I:peptide complexes.

EBV isolates from human populations with a high frequency of HLA A11 evade recognition by CTLs specific for an immunodominant A11-restricted epitope derived from the EBV nuclear antigen 4 (EBNA-4). We have previously described four nonimmunogenic variants of this epitope carrying single amino acid substitutions in the anchor residues of the peptide. We have now investigated the antigenicity, A11 binding capacity, endoplasmic reticulum translocation, endogenous processing, and presentation of these variants. The nonimmunogenic peptides were either unable to bind to HLA A11 or formed complexes of significantly lower stability compared with the immunogenic epitope. The latter peptides were produced in relatively large amounts by endogenous processing of EBNA-4 and associated with A11 molecules almost as efficiently as the immunogenic epitope, but the complexes failed to accumulate at the cell surface. The defect was not reversed by incubation of lymphoblastoid cell lines carrying the variant EBV strains at 26 degrees C. CTL lysis of HLA A11 positive targets was achieved by expressing one of the nonimmunogenic peptides through a vaccinia recombinant. However, the amount of peptide required for CTL sensitization exceeded, by at least 30-fold, that required for recognition of the immunogenic epitope. Collectively, these results suggest that complexes containing the nonimmunogenic peptides are formed but are then destroyed intracellularly. Thus, a specialized sorting mechanism seems to contribute in shaping the repertoire of peptides presented to T lymphocytes.

Cell Line, Transformed↗
PubMed1996

Defective presentation of MHC class I-restricted cytotoxic T-cell epitopes in Burkitt's lymphoma cells.

Defects of antigen processing/presentation have been suggested to play a role in the escape of Burkitt's lymphoma (BL) from cytotoxic T lymphocyte (CTL)-mediated rejection. Impaired presentation of an immunodominant HLA A11-restricted epitope from the resident or recombinant vaccinia virus-expressed Epstein-Barr virus nuclear antigen (EBNA)4 was demonstrated in the EBV-positive E95B-BL28 and its EBV-negative parental BL28 cell lines. We have investigated whether this was due to (i) impaired production of the antigenic peptide, (ii) poor peptide translocation into the ER lumen or (iii) inefficient maturation and transport of the MHC-peptide complexes at the cell surface. The defect was not overcome by cytosolic expression of a pre-formed epitope, suggesting that presentation of EBNA4 is not limited by inefficient production of the antigenic peptide. BL28 expressed 5- to 10-fold lower levels of the transporter associated with antigen presentation (TAP) 1 and TAP2 proteins and behaved poorly in a streptolysin-O-mediated peptide translocation assay, whereas E95B-BL28 showed higher TAP expression and virtually normal transporter function. Up-regulation of HLA A11 and reconstitution of TAP activity by treatment with IFN-gamma did not restore presentation of the resident EBNA4 in E95SB-BL28 and did not enhance presentation of the vaccinia virus-expressed intact protein or preformed epitope. Efficient maturation of class I molecules to Endo H-resistant species was demonstrated in pulse-chase experiments. Taken together, our findings identify a previously uncharacterized defect of antigen presentation which appears to affect events occurring after proteasomal degradation but before TAP-dependent peptide transport and MHC class I assembly and maturation.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1996

The life span of major histocompatibility complex-peptide complexes influences the efficiency of presentation and immunogenicity of two class I-restricted cytotoxic T lymphocyte epitopes in the Epstein-Barr virus nuclear antigen 4.

We have investigated the reactivity to two human histocompatibility leukocyte antigen (HLA) A11-restricted cytotoxic T lymphocyte (CTL) epitopes derived from amino acids 416-424 (IVTDFSVIK, designated IVT) and 399-408 (AVFDRKSVAK, designated AVF) of the Epstein-Barr virus (EBV) nuclear antigen (EBNA) 4. A strong predominance of CTL clones specific for the IVT epitope was demonstrated in polyclonal cultures generated by stimulation of lymphocytes from the EBV-seropositive donor BK with the autologous B95.8 virus-transformed lymphoblastoid cell line (LCL). This was not due to intrinsic differences of CTL efficiency since clones specific for the two epitopes lysed equally well A11-positive phytohemagglutinin blasts and LCLs pulsed with the relevant synthetic peptide. Irrespective of the endogenous levels of EBNA4 expression, untreated LCLs were lysed more efficiently by the IVT-specific effectors, suggesting that a higher density of A11-IVT complexes is presented at the cell surface. In accordance, 10-50-fold higher amounts of IVT peptides were found in high-performance liquid chromatography fractions of acid extracts corresponding to an abundance of about 350-12,800 IVT and 8-760 AVF molecules per cell. Peptide-mediated competition of CTL sensitization, transport assays in streptolysin-O permeabilized cells, and induction of A11 expression in the transporter associated with antigen presentation-deficient T2/A11 transfectant demonstrated that the IVT and AVF peptides bind with similar affinities to A11, are translocated with equal efficiency to the endoplasmic reticulum, and form complexes of comparable stability over a wide range of temperature and pH conditions. A rapid surface turnover of A11 molecules containing the AVF peptide was demonstrated in metabolically active T2/A11 cells corresponding to a half-life of approximately 3.5 as compared to approximately 2 h for molecules induced at 26 degrees C in the absence of exogenous peptides and >12 h for IVT-containing complexes. This difference in persistence is likely to determine the representation of individual class I-restricted CTL epitopes within the cell surface pool of molecules, and may be an important factor contributing to their immunogenicity.

Amino Acid Sequence↗
PubMed1996

HLA-A11-mediated protection from NK cell-mediated lysis: role of HLA-A11-presented peptides.

The capacity of MHC class I to protect target cells from NK is well established, but the mechanism by which these molecules influence NK recognition and the physical properties associated with this function remain poorly defined. We have examined this issue using as a model the HLA-A11 allele. HLA-A11 expression correlated with reduced susceptibility to NK and interferon-activated cytotoxicity in transfected sublines of the A11-defective Burkitt's lymphoma WW2-BL and the HLA class I A,B-null C1R cell line. Protection was also achieved by transfection of HLA-A11 in the peptide processing mutant T2 cells line (T2/A11), despite a very low expression of the transfected product at the cell surface. Induction of surface HLA-A11 by culture of T2/A11 cells at 26 degrees C or in the presence of beta 2m did not affect lysis, whereas NK sensitivity was restored by culture in the presence of HLA-All-binding synthetic peptides derived from viral or cellular proteins. Acid treatment rendered T2/A11 and C1R/A11 cells sensitive to lysis, but protection was restored after preincubation with peptide preparations derived from surface stripping of T2/A11 cells. Similar peptide preparations from T2 cells had no effect. The results suggest that NK protection is mediated by HLA-A11 molecules carrying a particular set of peptides that are translocated to the site of MHC class I assembly in the ER in a TAP-independent fashion.

Antigen Presentation↗
PubMed1996

Solvent exposed side chains of peptides bound to HLA A*1101 have similar effects on the reactivity of alloantibodies and specific TCR.

Peptides can affect the recognition of MHC class I molecules by allospecific antibodies. Two explanations have been proposed for this phenomenon. The 'conformational change' hypothesis suggests that peptide binding affects the availability of serologic determinants in the class I alpha1 and alpha2 domains while the 'peptide-side-chain effect' predicts that solvent exposed residues in the peptide are part of the serologic epitope. We have tested these possibilities by examining the recognition of peptide loaded HLA A*1101 molecules expressed in transporters associated with antigen processing (TAP)-deficient cell lines by three A11-specific mAb, and by comparing the effect of peptide analogues on the recognition of A11 complexes containing peptide epitopes from the Epstein-Barr virus nuclear antigen EBNA4 by antibodies and cytotoxic T lymphocytes (CT). The AUF5.13 and HB164 antibodies showed selective recognition of A11 molecules bound to partially overlapping sets of peptides from viral or cellular origin. The peptide dependence of AUF5.13 was confirmed in reconstitution experiments where A11 molecules were refolded at the surface of TAP-deficient T2/A11 cells that had been cultured at 26 degrees C and treated at pH3. Molecular modelling and Ala scanning mutagenesis of the IVTDFSVIK (IVT) and AVFDRKSDAK (AVF) peptides demonstrated that solvent-exposed peptide side chains affect CTL recognition as well as antibody binding. Substitution of Phe-P5 or Ser-P6 of the IVT peptide with Arg or Lys inhibited AUF5.13 recognition while binding was induced by substitution of the Arg-P5 and Lys-P6 of the AVF peptide with Ala. The results suggest that some allospecific antibodies recognized the surface of MHC class I-peptide complexes in a fashion similar to the TCR. This may involve direct interaction with the peptide side chains as well as recognition of peptide-induced perturbations in the class I complex.

ATP-Binding Cassette Transporters↗