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Most gamma delta T cells develop normally in the absence of MHC class II molecules.

MHC class I and class II molecules are essential for intrathymic maturation of a normal repertoire of alpha beta T cells. The development of gamma delta T cells may similarly require exposure to conventional MHC or MHC-like molecules for appropriate negative and positive selection. The availability of mice that are devoid of cell surface expression of MHC class II molecules allowed us to test directly the hypothesis that conventional class II molecules play a role in the development of gamma delta T cells. The proportions of gamma delta cells in thymus, lymph nodes, and spleens were indistinguishable between class II-deficient mice and littermate controls by FACS analysis. gamma delta T cells from class II-deficient mice proliferated normally in response to anti-TCR mAb. Examination of epidermal sheets from ear, torso, and tail skin demonstrated that class II-deficient mice had the same population density and distribution of gamma delta+ dendritic epidermal T cells as that of control littermates. gamma delta Cells in the epidermis of class II-deficient mice expressed Thy-1 and CD3 and were predominantly V gamma 3+. There were no significant differences in the immunophenotype of class II-deficient mice and their normal littermates in two-color immunofluorescence analysis of intestinal intraepithelial lymphocytes. Class II-deficient mice and control mice had 29 to 39% gamma delta bearing intestinal intraepithelial lymphocytes, of which 16 to 21% expressed V delta 4. Dendritic cells that stained positively for Thy-1, CD3, and gamma delta were identified in the vaginal epithelium of class II-deficient mice and their normal littermates. Our results indicate that MHC class II expression is not essential for the development of most gamma delta T cells.

Animals↗

Viral peptide specific induction of MHC class II expression by murine T cell clones.

We report that I-Ab-restricted T cell clones, elicited by influenza infection of C57BL/10 mice and specific for the hemagglutinin peptide HA1 186-205, express class II. They respond to peptide stimulation by IL release (IL-3 or IFN-gamma) without a requirement for APC but do not proliferate. Moreover, surface expression of class II requires de novo synthesis in the presence of the stimulatory peptide and is inhibited by coculture with TCR-specific Ab, or brefeldin A or cycloheximide. Clonotypic specificity of peptide induction was confirmed by failure of other allele specific peptides to enhance class II expression. Addition of the viral peptide to T cells induced homotypic adhesion, which provides a physical basis for stabilization of class II-peptide complexes at the cell surface. Extinction of class II expression was evident in the corresponding T cell hybridomas, which might account for the failure to report class II expression by murine T cells. Control studies indicated that class II was not passively acquired from APC by demonstrating 1) failure of processed Ag to induce class II expression, 2) allo-class II (Ak) was not acquired by coculture with peptide and semisyngeneic (H-2 b/k) APC, 3) absence of class II expression by a NP peptide-specific Th2 clone under identical culture conditions, and most significantly, 4) reverse-transcriptase PCR amplification and surface expression of class II using highly purified preparations of FACS-selected CD4+ class II- cells cocultured with the stimulatory peptide.

Alleles↗

Attenuation of HLA-DR expression by mononuclear phagocytes infected with Mycobacterium tuberculosis is related to intracellular sequestration of immature class II heterodimers.

MHC class II expression was examined in macrophages infected with Mycobacterium tuberculosis. IFN-gamma increased the surface expression of class II molecules in THP-1 cells and this was markedly reduced in cells infected with M. tuberculosis. Despite this effect, steady state levels of HLA-DRalpha, HLA-DRbeta, and invariant (Ii) chains were equivalent in control and infected cells. Metabolic labeling combined with pulse-chase experiments and biochemical analysis showed that the majority of class II molecules in infected cells became resistant to endoglycosidase H, consistent with normal Golgi processing. However, results of intracellular staining and dual color confocal microscopy revealed a significant defect in transport of newly synthesized class II molecules through the endocytic compartment. Thus, compared with findings in control cells, class II molecules in infected cells colocalized to a minimal extent with a lysosomal-associated membrane protein-1+ endosomal compartment. In addition, in contrast to control cells, class II molecules in infected cells failed to colocalize with endocytosed BSA under conditions where this marker is known to label late endosomes, lysosomes, and the MHC class II compartment. Consistent with defective transport along the endocytic pathway, the maturation of SDS-stable class II alphabeta dimers--dependent upon removal of Ii chain and peptide loading of class II dimers in the MHC class II compartment--was markedly impaired in M. tuberculosis-infected cells. These findings indicate that defective transport and processing of class II molecules through the endosomal/lysosomal system is responsible for diminished cell surface expression of MHC class II molecules in cells infected with M. tuberculosis.

Antigens, Differentiation, B-Lymphocyte↗

Reduced membrane major histocompatibility complex class I density and stability in a subset of human renal cell carcinomas with low TAP and LMP expression.

Suppression of MHC class I expression is thought to allow tumor cells to escape immune surveillance mediated by CD8(+) CTLs. For stable MHC class I surface expression, multiple protein interactions are required for efficient assembly of MHC class I heavy chain and beta 2-microglobulin with endogenous peptides. Peptide processing and transport into the endoplasmic reticulum involves the genes of the transporters associated with antigen processing, TAP-1 and TAP-2, and the two components of the proteasome complex, the low molecular weight proteins LMP-2 and LMP-7. We selected human renal cell carcinoma (RCC) cells derived from a tumor that is thought to be controlled by host immunity to study the MHC class I antigen presentation machinery. Eleven RCC lines established from primary tumors were investigated for the mRNA and protein expression of MHC class I, TAP, and LMP genes. In addition, membrane stability of MHC class I was determined by incubation of the RCC cell lines at low temperature and in the presence of exogenous HLA-binding peptides. Our results revealed the existence of two different phenotypes of RCC cell lines. Group I displayed temperature-stable MHC class I surface expression associated with high, and in most cases coordinated, expression of MHC class I heavy and light chain, TAP and LMP transcripts, and proteins. Group II demonstrated a marked MHC class I instability at 37 degreesC associated with low but coordinated expression of the respective transcripts and proteins. MHC class I membrane expression of group II, but not of group I RCC cells, could be stabilized by incubation with specific MHC class I binding peptides. These results suggest an important role of the genes of the antigen presentation machinery in stable and efficient MHC class I surface expression of RCC cells. However, it has still to be defined whether deficient antigen processing is one of the mechanisms of RCC cells to escape the surveillance of the immune system.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

The structure of posttraumatic stress disorder: latent class analysis in 2 community samples.

CONTEXT: Latent structure analysis of DSM-IV posttraumatic stress disorder (PTSD) can help clarify how persons who experience traumatic events might be sorted into clusters with respect to their symptom profiles. Classification of persons exposed to traumatic events into clinically homogeneous groups would facilitate further etiologic and treatment research, as well as research on the relationship of trauma and PTSD with other disorders. OBJECTIVES: To examine empirically the structure underlying PTSD criterion symptoms and identify discrete classes with similar symptom profiles. DESIGN: Data on PTSD symptoms from trauma-exposed subsets of 2 community samples were subjected to latent class analysis. The resultant classes were studied in associations with trauma type and indicators of impairment. SETTING: The first sample is from the Detroit Area Survey of Trauma (1899 trauma-exposed respondents with complete data) and the second is from a mid-Atlantic study of young adults conducted by The Johns Hopkins University Prevention Research Center, Baltimore, Md (1377 trauma-exposed respondents with complete data). PARTICIPANTS: Respondents in the 2 community samples who experienced 1 or more qualifying PTSD-level traumatic events. MAIN OUTCOME MEASURES: Number, size, and symptom profiles of latent classes. RESULTS: In both samples, analysis yielded 3 classes: no disturbance, intermediate disturbance, and pervasive disturbance. The classes also varied qualitatively, with emotional numbing distinguishing the class of pervasive disturbance, a class that approximates the subset with DSM-IV PTSD. Members of the pervasive disturbance class were far more likely to report use of medical care and disruptions in life or activities. CONCLUSIONS: The 3-class structure separates trauma-exposed persons with pervasive disturbance (a class that approximates DSM-IV PTSD) from no disturbance and intermediate disturbance, a distinction that also helps identify population subgroups with low risk for any posttrauma disturbance. The results suggest that the structure of PTSD is ordinal and configurational and that emotional numbing differentiates the class with pervasive disturbance. These results should motivate prospective research of persons who have experienced trauma to trace the emergence of posttrauma symptoms and the timing of emotional numbing relative to other symptoms.

Adult↗

Clinical and survival covariates of eight classes of childhood supratentorial neuroglial tumors.

BACKGROUND: In the current study, the authors investigated clinical, surgical, and histologic characteristics (covariates) and their interactions in eight previously identified classes of childhood supratentorial neuroglial tumors. The classes resulted from 5 factor score profiles on 703 supratentorial neuroglial tumors in the Childhood Brain Tumor Consortium database. METHODS: The Cox proportional models were used to identify class survival covariates. RESULTS: Age was found to be a survival covariate only in Class 1, in which older age increased the 5-year survival rate 73% from the first year (0.49) to the tenth year (0.85). A greater amount of tumor removed improved survival in Classes 2 and 4 only. Rosenthal fibers improved survival in Class 2 and overrode the negative effects of high Proliferative factor scores and pleomorphic nuclei. Survival for Class 3 children with high Proliferative factor scores improved from 0.60 to 0.95 as the Spongy factor scores increased. Survival in Class 4 increased from 0.17 to 0.39 with total tumor removal. Irregular nuclei and glomeruloid capillaries improved survival in Class 5 patients. Class 6 survival improved with low cell density. Macrocysts in tumors in Classes 1 and 5 were found to improve survival. CONCLUSIONS: As a result of the current study, the authors conclude that survival covariates differ with tumor class and may modify prognosis considerably.

Astrocytoma↗

Characterization of three CpG oligodeoxynucleotide classes with distinct immunostimulatory activities.

Oligodeoxynucleotides (ODN) with unmethylated deoxycytidyl-deoxyguanosine (CpG) dinucleotides (CpG ODN) mimic the immunostimulatory activity of bacterial DNA and are recognized by the Toll-like receptor 9 (TLR9). CpG ODN of the B-Class stimulate strong B cell and NK cell activation and cytokine production. The highest degrees of NK stimulation as well as IFN-alpha secretion by plasmacytoid DC were found to occur only with A-Class ODN. A third class of CpG ODN combines the immune effects of A- and B-Class CpG ODN. C-Class ODN strongly stimulate B cell or NK cell activation and IFN-alpha production. In contrast to the A-Class, the C-Class is wholly phosphorothioate, has no poly-G stretches, but has palindromic sequences combined with stimulatory CpG motifs. All classes stimulate TLR9-dependent signaling, but with strikingly different dose-response relationships that are quite in contrast to those observed for IFN-alpha. Effects similar to those on human cells were observed on mouse splenocytes. In contrast, splenocytes from TLR9-deficient mice did not show any response to the three CpG ODN classes. In vivo studies demonstrate that C-Class ODN are very potent Th1 adjuvants. C-Class ODN may represent new therapeutic drugs that combine the effects of A- and B-Class ODN for broad applications in infectious disease or cancer therapy.

Animals↗

Detection of IgA class circulating immune complexes bound to anti-C3d antibody in patients with IgA nephropathy.

IgA class circulating immune complexes (CIC) were detected by solid-phase fluorescent enzyme immunoassay of F(ab')2 anti-C3d antibody in the serum of 52 patients with IgA nephropathy. Conglutinin (Kg) binding IgA class CIC were also measured, and results by these assays were compared. Kg binding IgA class CIC and anti-C3d binding IgA class CIC were detected in 27% and 44%, respectively, of the patients with IgA nephropathy. Either or both of the two were found in 65% of the patients. There was no significant correlation between IgA class CIC detected by these methods and serum IgA. Although all samples with a very high level of anti-C3d binding IgA class CIC did not also have a very high level of Kg binding IgA class CIC, there was a slight quantitative correlation between the 2 assays. Ultracentrifugation analysis showed that anti-C3d binding IgA class CIC were of various sizes between polymeric (21 S) and monomeric IgA (7 S), whereas Kg binding IgA class CIC were mostly monomeric IgA (8 S) with a minor component of heavy fractions (14 S). Both IgA class CIC fixed iC3b and IgA class CIC fixed C3d are present in IgA nephropathy. These observations suggest that the different types of complement bound to IgA class CIC have different roles in IgA nephropathy.

Antigen-Antibody Complex↗

Enhanced expression of HLA-class II molecules on activated human T lymphocytes following treatment with tumor necrosis factor alpha.

Many factors induce or enhance expression of major histocompatibility complex class I and class II molecules on various cell types. Human T lymphocytes are class II negative in the resting state but show expression of class II molecules following activation. We analyzed the modulating capacity of the lymphokines recombinant interferon gamma (rIFN-gamma), interleukin-4 (IL-4), and recombinant tumor necrosis factor alpha (rTNF-alpha) on class II expression in subsets of alloactivated human T lymphocytes. The activated CD4+ T cells expressed all three class II isotypes (DR, DQ, and DP), whereas the cytotoxic CD8+ T-cell lines expressed DR and DP molecules but failed to bind DQ-specific monoclonal antibodies significantly. Treatment with rIFN-gamma and IL-4 had no effect on class II expression on any of the T-cell lines or clones, whereas rTNF-alpha enhanced class II expression in both subsets. rTNF-alpha could modulate expression of all three class II isotypes but, in principle, it appears only to affect ongoing class II synthesis as de novo synthesis of class II molecules with a resultant change in the class II phenotype from DR+ DQ- DP+ to DR+ DQ+ DP+ in the CD8+ T lymphocytes was not observed. No synergic effects of rINF-gamma and rTNF-alpha were observed; this results from the fact that activated T cells express few, if any, receptors of rIFN-gamma.

Antibodies, Monoclonal↗

C-Class CpG ODN: sequence requirements and characterization of immunostimulatory activities on mRNA level.

Synthetic oligodeoxynucleotides (ODN) containing unmethylated deoxycytosine-deoxyguanosine (CpG) motifs are very potent inducers of the innate immune system, mimicking the effects of bacterial DNA. CpG ODN are recognized by Toll-like receptor 9 (TLR9). Three classes of TLR9 agonists have been described: B-Class CpG ODN that induce strong B- and NK-cell activation and A-Class ODN that induce very high levels of IFN-alpha by plasmacytoid dendritic cells. The recently described C-Class ODN combine most efficiently properties of A- and B-Class ODN in that they induce strong B-cell activation comparable to B-Class ODN together with IFN-alpha secretion comparable to A-Class ODN. Here, we investigate sequence requirements of C-Class ODN regarding optimal IFN-alpha secretion. Sequence as well as backbone modifications like 2'-O-methyl modifications especially in the 5' part of the ODN influence IFN-alpha-producing capacity. Kinetic studies on mRNA level for CD69, IFN-gamma, IP-10 and IL-18 by semi-quantitative PCR demonstrated differences in mRNA transcription for some cytokines suggesting different regulatory mechanisms for different ODN classes. High amounts of IP-10 mRNA and protein as well as up-regulation of IL-18 mRNA were observed especially for the A- and C-Classes. According to these data, C-Class ODN can be described as strong Th1 inducers with the stimulation of type I and II interferon as well as IP-10 production and strong NK activation. These characteristics can be availed to induce potent anti-tumor or anti-viral effects. Consequently, C-Class CpG ODN represent ideal drug candidates for anti-viral and/or anti-tumor therapy.

Antigens, CD↗

Patients with brain metastases: hope for recursive partitioning analysis (RPA) class 3.

PURPOSE: The objectives of the present study were (a) to validate the prognostic classification derived from recursive partitioning analysis (RPA) of the Radiation Therapy Oncology Group (RTOG); (b) to identify prognostic factors in class 3; (c) to examine the impact of treatment related variables on the prognosis in class 3. PATIENTS AND METHODS: Nine hundred and sixteen patients with brain metastases had resection and whole brain radiotherapy (WBRT, n = 257) or WBRT alone (n = 659) at our institution from 1985 to 2000. Patients were grouped into RPA classes 1, 2, and 3 (n = 67, 441, and 408, respectively). RESULTS: Median survival of the whole group was 3.4 months. Median survival in classes 1, 2, and 3 was 8.2, 4.9, and 1.8 months, respectively. In class 3, age (<65 years vs. > or =65 years, relative risk (RR) 0.75), status of the primary tumor (controlled vs. uncontrolled, RR 0.86), and the number of brain metastases (single vs. multiple, RR 0.76) were independent prognostic variables. We defined three prognostic subgroups: class 3a (n = 51): age <65 years, controlled primary tumor, single brain metastasis; class 3c (n = 44): age > or =65 years, uncontrolled primary tumor, multiple brain metastases; class 3b (n = 313): all other patients. Median survival in classes 3a, 3b, and 3c was 3.2, 1.9, and 1.2 months, respectively (P < 0.0001). Intra-class comparisons showed that resection followed by WBRT yielded significantly better survival compared with WBRT alone. CONCLUSION: Our results validate the RTOG RPA classification for patients with brain metastases. The variables age, status of the primary, and number of brain metastases allow the division of class 3 into prognostic subgroups. Even class 3 patients may benefit from more aggressive treatment strategies.

Adult↗

Activity of class I, II, III, and IV alcohol dehydrogenase isoenzymes in human gastric mucosa.

The gastric alcohol dehydrogenase (ADH) plays an important role in the "first pass" metabolism of ethanol. Human ADH exists in multiple forms, grouped into six classes and located in different tissues. In present study we investigated the activity of four classes (I, II, III, and IV) of alcohol dehydrogenase isoenzymes in the different parts of stomach (corpus and antrum) in patients with suspected gastric ulcer. The aim of the study was assess the particular role of different classes of ADH in the gastric dehydrogenase activity. For the measurement of the activity of class I and class II isoenzymes, we employed new fluorometric methods with specific substrates. The activity of class III alcohol dehydrogenase was measured by the photometric method with n-octanol and class IV with m-nitrobenzaldehyde as a substrate, respectively. All biopsy specimens were taken from less changed areas of the antrum and body of the stomach of 68 patients suspected of having gastric ulcer. It was found that ADH IV (gastric) activity was the highest (14.76 +/- 0.68 in the corpus of the stomach in men; and 7.61 +/- 0.68 in women, respectively). The activity of the ADH III isoenzyme was lower than that of ADH IV. The activities of class I and II ADH isoenzymes were barely detectable. All tested classes of ADH had higher activity in the corpus than in the antrum and in males than in females. In conclusion, the most important form of gastric ADH is isoenzyme of class IV, less important is the isoenzyme of class III. ADH classes I and II seem to have no role in the stomach.

Adult↗

Carvedilol improves functional class in patients with severe left ventricular dysfunction referred for heart transplantation.

PURPOSE: Limited options are available to improve the functional class of patients awaiting cardiac transplantation. We assessed the effect of carvedilol on New York Heart Association (NYHA) class, heart rate (HR), blood pressure (BP), jugular venous pressure (JVP), electrolytes and renal function in patients with markedly decreased left ventricular (LV) function referred for cardiac transplantation assessment. METHODS: Sixty-one patients (age = 52 +/- 12 yr, EF = 23 +/- 7%, VO2 max = 16 +/- 5 mL/kg/min) referred to the cardiac transplant clinic were reviewed before and after the addition of carvedilol (starting dose 3.125 mg twice daily to target dose of 25 mg twice daily) to usual heart-failure therapy. Over a 1-yr period, at each visit prior to initiation, at baseline initiation visit and at each follow-up visit, NYHA class, BP, HR, JVP, electrolytes, and renal function were obtained. Statistical analysis was performed using random effects regression approach. A multiple logistic regression analysis was performed on 52/61 patients to determine possible associations between NYHA improvement and the following patient characteristics: sex, etiology of cardiomyopathy, initial NYHA class, and dose of carvedilol. RESULTS: Three patients died (2 after stopping carvedilol) and 3 were transplanted. Median follow-up was 100 d. Sixteen patients reached the target dose after a mean of 137 d (2.75 visits). Estimated time-to-target dose is 8 months (5.6 visits). BP tended to increase (p = 0.07 for change in trend) with no change in JVP, electrolytes or renal function. HR decreased 6 +/- 3 b.p.m. (p = 0.03). Of 14 patients NYHA class I/II at baseline, none were class III/IV at last follow-up visit. Of 47 patients NYHA class III/IV at baseline, 25 were class I/II, and 22 were class III/IV at last follow-up (p < 0.001). Multiple logistic regression analysis did not demonstrate any factor predictive of subsequent NYHA class improvement. CONCLUSIONS: Despite less than target doses in most patients, a favorable effect of carvedilol on functional class in patients with severe congestive heart failure (CHF) referred for transplant was observed. Those with good functional status remained stable and those with poor functional status improved or remained stable. No baseline patient characteristic predicted improvement. The shortage of donor organ requires optimal use of medical regimens which may improve functional class while awaiting transplantation and which may delay the necessity for heart transplantation. Therefore, addition of carvedilol to usual medical therapy may be beneficial even in patients with severe LV dysfunction and poor NYHA classification.

Adrenergic beta-Antagonists↗

Reappraisal of Class II molar relationships diagnosed from the lingual side.

The buccal and lingual molar relationships in 459 clinically diagnosed Class II cases and the impact of upper first permanent molar rotation on this classification were assessed. Of the 459 Class II cases, 99 (10.78%) sides exhibited unilateral neutral molar relationship assessed from both the buccal and the lingual aspect. None of the molars exhibiting Class I relationship were rotated. The 175 (19.06%) sides with mild Class II molar relationship from the buccal aspect were all judged to be Class I lingually and 85% of these molars were rotated mesially. In the case of a moderate buccal Class II tendency, 55% of the molars were in a Class I relationship when evaluated from the lingual side, whereas 45% belonged to mild (1st degrees ) Class II with molar rotations present in 81% of the cases. When a severe (3rd degrees ) Class II was present from the buccal side, its corresponding lingual Class II tendencies were accordingly elevated (83% 2nd degrees, 17% 3rd degrees ). Among these cases, the molar rotations occurred in 74% of the cases. The results suggested the upper mesiopalatal cusp as a pivotal axis around which the upper first molar rotated. This cusp occluded most frequently in the central fossa, less frequently in the mesio-triangular fossa and rarely anterior to the mesio-triangular fossa. Rotations that correspond to these types of lingual occlusion were 85, 81 and 74%, respectively. The correlation between buccal and lingual molar relationships and their correlation with molar rotation were all significant (p<0.001). Moreover, canine relationship was linearly correlated with the conventional (buccal) molar relationship in Class II cases, expressed by Ycanine relation=1.423+0.47 Xmolar relation (p<0.001). The results indicate that a more differentiated diagnosis of the molar relationship will allow for a more causally directed correction of Class II molar relationship.

Journal Article↗

High resolution HLA class I and II typing and CTLp frequency in unrelated donor transplantation: a single-institution retrospective study of 69 BMTs.

UNLABELLED: The results of unrelated donor transplantation (URD-BMT) are difficult to analyze since the continuous advances in HLA typing technology allow the detection of new mismatches unknown at the time of transplantation. We sought to confirm that matched recipient-donor pairs are in fact often mismatched when advanced HLA typing techniques are used. We retrospectively studied the impact of the results of high resolution HLA typing for HLA class I (-A, -B, -C) and HLA class II (-DR, -DQ, -DP) loci, and cytotoxic T lymphocyte precursor (CTLp) frequency, on the outcome of 69 URD-BMT procedures. At the time of transplant, six (6/69) and two (2/69) donor-recipient pairs were mismatched for HLA class I (-A and -B by serology) and HLA class II, respectively, while one pair was mismatched for both HLA class I and II. Using high resolution DNA typing, HLA class I mismatches were found in 31 (45%) pairs and HLA class II mismatches in nine (13%) pairs. Twenty-three of the 69 pairs were HLA-C mismatched. Low CTLp frequencies were found among the 19 HLA class I matched pairs tested, and also in 5/14 mismatched pairs (of whom three had severe aGVHD). The overall survival of the cohort was 28 +/- 6%. Among the 33 patients who were fully matched with their donors, the survival rate was 66% in the 18 patients with a standard hematological risk and 9% in the 15 high risk patients. Only two of the 33 patients developed severe aGVHD, and only one had graft rejection. Among the 36 mismatched pairs, the survival rate was 31% in the 13 patients with a standard hematological risk and 8% in the 23 high risk patients. Sixteen of these 36 patients died from severe aGVHD and four had graft failure or rejection. Three of the 10 patients with only an HLA-C mismatch died from severe aGVHD, and two had graft rejection. IN CONCLUSION: (1) donor-recipient matching based on high resolution HLA class I and II DNA typing is associated with significantly better outcome after URD-BMT; (2) the results of URD-BMT with classical GVHD prevention are comparable to those of geno-identical BMT when donor and recipient are fully matched for HLA-A, -B, -C, -DRB1 and -DQB1 on the basis of high resolution typing; (3) CTLp frequencies do not correlate constantly with HLA class I matching, and our results fail to show that CTLp assay can distinguish between permissible and non-permissible class I mismatches; (4) clinical trials involving donor-recipient pairs with known HLA class I mismatches are needed to improve aGVHD prevention without increasing graft failure rate.

Adult↗

Expression of MHC class II in T cells is associated with increased HIV-1 expression.

HIV-1 replicates in activated T cells at significantly higher levels than in resting cells. Thus, certain molecules up-regulated during T cell activation appear to be important for HIV-1 replication. In this study, we present evidence suggesting that expression of MHC class II (class II) molecules on CD4+ T cells facilitate HIV-1 replication. T cells that expressed class II supported greater virus replication than T cells lacking class II. The class II+ cells, when either infected with HIV-1 or transfected with an env-minus HIV-1 provirus plasmid, produced 10-20-fold greater virus expression than class II- cells. Anti-class II antibody markedly inhibited virus expression in class II+ cells (but not class II- cells) and also decreased the nuclear binding activity of AP-1, an inducible transcription factor important in T cell activation and HIV-1 expression. Most importantly, the induction of class II expression by transfection of the MHC class II transactivator (CIITA) stimulated HIV-1 replication in Jurkat T cells. Taken together, these data suggest that expression of MHC class II molecules and/or CIITA in T cells enhances HIV-1 transcription.

CD4-Positive T-Lymphocytes↗

Distribution of class I, III and IV alcohol dehydrogenase mRNAs in the adult rat, mouse and human brain.

The localization of different classes of alcohol dehydrogenases (ADH) in the brain is of great interest because of their role in both ethanol and retinoic acid metabolism. Conflicting data have been reported in the literature. By Northern blot and enzyme activity analyses only class III ADH has been detected in adult brain specimens, while results from riboprobe in situ hybridization indicate class I as well as class IV ADH expression in different regions of the rat brain. Here we have studied the expression patterns of three ADH classes in adult rat, mouse and human tissues using radioactive oligonucleotide in situ hybridization. Specificity of probes was tested on liver and stomach control tissue, as well as tissue from class IV ADH knock-out mice. Only class III ADH mRNA was found to be expressed in brain tissue of all three investigated species. Particularly high expression levels were found in neurons of the red nucleus in human tissue, while cortical neurons, pyramidal and granule cells of the hippocampus and dopamine neurons of substantia nigra showed moderate expression levels. Purkinje cells of cerebellum were positive for class III ADH mRNA in all species investigated, whereas granular layer neurons were positive only in rodents. The choroid plexus was highly positive for class III ADH, while no specific signal for class I or class IV ADH was detected. Our results thus support the notion that the only ADH expressed in adult mouse, rat and human brain is class III ADH.

Aged↗

Regulation of genes for HLA class II antigens in cell lines from patients with severe combined immunodeficiency.

HLA Class II-negative severe combined immunodeficiency (SCID) results from a congenital defect characterized by an absence of HLA Class II antigens. Patients with the disorder have no HLA-DR, DQ, or DP antigens or mRNAs in their peripheral-blood lymphocytes. The affected gene is a recessive, transacting regulatory gene that controls the expression of Class II genes. We studied the regulation of HLA Class II gene expression with the use of established Epstein-Barr virus-transformed B-cell lines and skin fibroblast lines from a group of patients with SCID. Lymphoblastoid B-cell lines from the patients contained no mRNA for HLA-DR, DQ, and DP alpha and beta polypeptides, but did express mRNA for the HLA-associated invariant chain, which is normally coregulated with HLA Class II antigens. In the B-cell line from one patient, a very low amount of DR mRNA could be detected, indicating some heterogeneity in SCID. The lymphokine gamma-interferon, a strong inducer of Class II genes in a variety of normal cells, did not restore Class II gene expression in any of the SCID B-cell lines. More important, gamma-interferon was unable to induce any Class II mRNA in fibroblast lines from patients with SCID, in contrast to the efficient induction observed in normal fibroblasts. The invariant-chain gene, however, was induced in the SCID fibroblasts, confirming a unique uncoupling in the regulation of invariant and Class II genes. Thus, the genetic defect in patients with SCID affects not only the B-cell lineage but also the inducible expression of HLA Class II genes that is normally observed in Class II-negative cells, such as fibroblasts. This unresponsiveness to gamma-interferon in vitro indicates that patients with SCID will not respond to treatment with this lymphokine. Our data also increase understanding of the normal mechanisms regulating the genes for the HLA Class II cell-surface glycoproteins.

Cell Line↗