Transcriptional regulation of the invariant chain associated with MHC class II molecules

The invariant chain associated with the major histocompatibility complex (MHC) class II molecules is a non-polymorphic glycoprotein implicated in antigen processing and class II molecule intracellular transport. Class II molecules and invariant chain (In) are expressed primarily by B lymphocytes and antigen-presenting cells such as macrophages and can be induced by interferon gamma (IFN-$\gamma$) in a variety of cell types such as endothelial cells, fibroblasts, and astrocytes. In this study the cis-acting sequences involved in the constitutive, tissue-specific, and IFN-$\gamma$ induced expression of the human In gene were investigated and nuclear proteins which specifically bound these sequences were identified.^ To define promoter sequences involved in the regulation of the human In gene, 790 bp 5$\sp\prime$ to the initiation of transcription were subcloned upstream of the gene encoding chloramphenicol acetyl transferase (CAT). Transfection of this construct into In expressing and non-expressing cell lines demonstrated that this 790 bp In promoter sequence conferred tissue specificity to the CAT gene. Deletion mutants were created in the promoter to identify sequences important for transcription. Three regulatory regions were identified $-$396 to $-$241, $-$241 to $-$216, and $-$216 to $-$165 bp 5$\sp\prime$ to the cap site. Transfection into a human glioblastoma cell line, U-373 MG, and treatment with IFN-$\gamma$, demonstrated that this 5$\sp\prime$ region is responsive to IFN-$\gamma$. An IFN-$\gamma$ response element was sublocalized to the region $-$120 to $-$61 bp. This region contains homology to the interferon-stimulated response element (ISRE) identified in other IFN responsive genes. IFN-$\gamma$ induces a sequence-specific DNA binding factor which binds to an oligonucleotide corresponding to $-$107 to $-$79 bp of the In promoter. This factor also binds to an oligonucleotide corresponding to $-$91 to $-$62 of the interferon-$\beta$ gene promoter, suggesting this factor may be member of the IRF-1/ISGF2, IRF-2, ICSBP family of ISRE binding proteins. A transcriptional enhancer was identified in the first intron of the In gene. This element, located in a 2.6 kb BamHI/PstI fragment, enhances the IFN-$\gamma$ response of the promoter in U-373 MG. The majority of the In enhancer activity was sublocalized to a 550 bp region $\sim$1.6 kb downstream of the In transcriptional start site. ^

HLA class II antigen expression in colorectal carcinoma tumors as a favorable prognostic marker.

The goal of this study was to determine the frequency of HLA class II antigen expression in colorectal carcinoma (CRC) tumors, its association with the clinical course of the disease, and the underlying mechanism(s). Two tissue microarrays constructed with 220 and 778 CRC tumors were stained with HLA-DR, DQ, and DP antigen-specific monoclonal antibody LGII-612.14, using the immunoperoxidase staining technique. The immunohistochemical staining results were correlated with the clinical course of the disease. The functional role of HLA class II antigens expressed on CRC cells was analyzed by investigating their in vitro interactions with immune cells. HLA class II antigens were expressed in about 25% of the 220 and 21% of the 778 tumors analyzed with an overall frequency of 23%. HLA class II antigens were detected in 19% of colorectal adenomas. Importantly, the percentage of stained cells and the staining intensity were significantly lower than those detected in CRC tumors. However, HLA class II antigen staining was weakly detected only in 5.4% of 37 normal mucosa tissues. HLA class II antigen expression was associated with a favorable clinical course of the disease. In vitro stimulation with interferon gamma (IFNγ) induced HLA class II antigen expression on two of the four CRC cell lines tested. HLA class II antigen expression on CRC cells triggered interleukin-1β (IL-1β) production by resting monocytes. HLA class II antigen expression in CRC tumors is a favorable prognostic marker. This association may reflect stimulation of IL-1β production by monocytes.

Long-term evaluation of Class II subdivision treatment with unilateral maxillary first molar extraction

OBJECTIVE To evaluate the long-term effects of asymmetrical maxillary first molar (M1) extraction in Class II subdivision treatment. MATERIALS AND METHODS Records of 20 Class II subdivision whites (7 boys, 13 girls; mean age, 13.0 years; SD, 1.7 years) consecutively treated with the Begg technique and M1 extraction, and 15 untreated asymmetrical Class II adolescents (4 boys, 11 girls; mean age, 12.2 years; SD, 1.3 years) were examined in this study. Cephalometric analysis and PAR assessment were carried out before treatment (T1), after treatment (T2), and on average 2.5 years posttreatment (T3) for the treatment group, and at similar time points and average follow-up of 1.8 years for the controls. RESULTS The adjusted analysis indicated that the maxillary incisors were 2.3 mm more retracted in relation to A-Pog between T1 and T3 (β  =  2.31; 95% CI; 0.76, 3.87), whereas the mandibular incisors were 1.3 mm more protracted (β  =  1.34; 95% CI; 0.09, 2.59), and 5.9° more proclined to the mandibular plane (β  =  5.92; 95% CI; 1.43, 10.41) compared with controls. The lower lip appeared 1.4 mm more protrusive relative to the subnasale-soft tissue-Pog line throughout the observation period in the treated adolescents (β  =  1.43; 95% CI; 0.18, 2.67). There was a significant PAR score reduction over the entire follow-up period in the molar extraction group (β  =  -6.73; 95% CI; -10.7, -2.7). At T2, 65% of the subjects had maxillary midlines perfectly aligned with the face. CONCLUSIONS Unilateral M1 extraction in asymmetrical Class II cases may lead to favorable occlusal outcomes in the long term without harming the midline esthetics and soft tissue profile.

Antibodies against major histocompatibility complex class II antigens directly inhibit the growth of T cells infected with Theileria parva without affecting their state of activation

We have analyzed the effect of antibodies (Abs) directed against major histocompatibility complex (MHC) class II Abs on the proliferation of Theileria parva-infected (Tpi) T cells. Anti-MHC class II Abs exert a direct effect on Tpi T cells causing an acute block in their proliferation. The inhibition does not involve apoptosis and is also entirely reversible. The rapid arrest of DNA synthesis caused by anti-MHC class II Abs is not due to interference with the state of activation of the T cells since the transcriptional activator NF-kappa B remains activated in arrested cells. In addition, interleukin 2 (IL-2), IL-2R, and c-myc gene expression are also unaffected. By analyzing the cell-cycle phase distribution of inhibited cells, it could be shown that cells in all phases of the cell cycle are inhibited. The signal transduction pathway that results in inhibition was shown to be independent of protein kinase C and extracellular Ca2+. Tyrosine kinase inhibitors, however, partly reduced the level of inhibition and, conversely, phosphatase inhibitors enhanced it. The possible relevance of this phenomenon in other systems is discussed.

Avaliação de assimetria nas arcadas dentárias de indivíduos com oclusão normal e má oclusão de Classe II

A assimetria das arcadas dentárias constitui um assunto de grande importância estudado por profissionais de Ortodontia na elaboração de um diagnóstico correto, planejamento e execução de um tratamento ortodôntico bem sucedido. O objetivo deste estudo foi avaliar o grau de assimetria das arcadas dentárias em indivíduos com oclusão normal e má oclusão de Classe II, divisão 1 e 2, bem como o dimorfismo sexual existente. Foram avaliados 180 pares de modelos de estudo de indivíduos do sexo masculino e feminino, na faixa etária de 12 a 21 anos, divididos em 3 grupos de 60 pares de modelos, de acordo com a má oclusão. Os grupos foram classificados em: Grupo 1 - indivíduos com oclusão normal, Grupo 2 - indivíduos com má oclusão de Classe II divisão 1 (Cl II 1), e Grupo 3 - indivíduos com má oclusão de Classe II divisão 2 (Cl II 2). Os modelos foram medidos utilizando-se um aparelho inédito, idealizado e fabricado exclusivamente para a análise de assimetria das arcadas dentárias. Para a análise de assimetria foram realizadas 2 medições angulares desvio de linha média (DLM), posicionamento dos caninos (PC) e 3 lineares distância dos caninos à rafe palatina (DC), distância inter-caninos (DIC), posicionamento dos primeiros molares no sentido ântero-posterior (PM). Concluiu-se que a ocorrência de assimetria nas arcadas dentárias independe da má oclusão. O Grupo 1 apresentou um menor grau de assimetria nas arcadas dentárias em relação aos grupos 2 e 3, os quais apresentaram um grau de assimetria semelhante. O grau de assimetria nas arcadas dentárias inferiores, nos 3 grupos, foi maior do que nas arcadas dentárias superiores. A direção do desvio da linha média apresentou uma correlação inversamente proporcional do lado em que o molar se apresentava mesializado, nas arcadas superior e inferior dos três grupos, com exceção da arcada superior no Grupo 2 (Classe II divisão 1). Houve dimorfismo sexual estatisticamente significante para algumas medidas, porém é importante considerar os baixos valores e a disposição, destas diferenças, entre as medidas realizadas, a qual revela ter se tratado de dados obtidos ao acaso.

Avaliação de assimetria nas arcadas dentárias de indivíduos com oclusão normal e má oclusão de Classe II

A assimetria das arcadas dentárias constitui um assunto de grande importância estudado por profissionais de Ortodontia na elaboração de um diagnóstico correto, planejamento e execução de um tratamento ortodôntico bem sucedido. O objetivo deste estudo foi avaliar o grau de assimetria das arcadas dentárias em indivíduos com oclusão normal e má oclusão de Classe II, divisão 1 e 2, bem como o dimorfismo sexual existente. Foram avaliados 180 pares de modelos de estudo de indivíduos do sexo masculino e feminino, na faixa etária de 12 a 21 anos, divididos em 3 grupos de 60 pares de modelos, de acordo com a má oclusão. Os grupos foram classificados em: Grupo 1 - indivíduos com oclusão normal, Grupo 2 - indivíduos com má oclusão de Classe II divisão 1 (Cl II 1), e Grupo 3 - indivíduos com má oclusão de Classe II divisão 2 (Cl II 2). Os modelos foram medidos utilizando-se um aparelho inédito, idealizado e fabricado exclusivamente para a análise de assimetria das arcadas dentárias. Para a análise de assimetria foram realizadas 2 medições angulares desvio de linha média (DLM), posicionamento dos caninos (PC) e 3 lineares distância dos caninos à rafe palatina (DC), distância inter-caninos (DIC), posicionamento dos primeiros molares no sentido ântero-posterior (PM). Concluiu-se que a ocorrência de assimetria nas arcadas dentárias independe da má oclusão. O Grupo 1 apresentou um menor grau de assimetria nas arcadas dentárias em relação aos grupos 2 e 3, os quais apresentaram um grau de assimetria semelhante. O grau de assimetria nas arcadas dentárias inferiores, nos 3 grupos, foi maior do que nas arcadas dentárias superiores. A direção do desvio da linha média apresentou uma correlação inversamente proporcional do lado em que o molar se apresentava mesializado, nas arcadas superior e inferior dos três grupos, com exceção da arcada superior no Grupo 2 (Classe II divisão 1). Houve dimorfismo sexual estatisticamente significante para algumas medidas, porém é importante considerar os baixos valores e a disposição, destas diferenças, entre as medidas realizadas, a qual revela ter se tratado de dados obtidos ao acaso.

AVALIAÇÃO COMPARATIVA DA AGRADABILIDADE FACIAL DE PACIENTES COM MÁ-OCLUSÃO DE CLASSE II TRATADOS COM COMPENSAÇÃO DENTÁRIA OU CIRURGIA ORTOGNÁTICA

O objetivo deste estudo foi avaliar, por meio de fotografias em norma frontal e lateral, a agradabilidade facial obtida com o tratamento de pacientes portadores de má oclusão de classe II. Foram selecionados dois grupos de pacientes que receberam abordagens diferentes de tratamento, um submetido à cirurgia ortognática e o outro à compensação dentária. As fotografias em norma lateral e frontal obtidas ao início e final do tratamento foram distribuídas aleatoriamente e dispostas em apresentação de multimídia para serem submetidas à avaliação subjetiva de indivíduos leigos e ortodontistas, em uma escala linear crescente. Os resultados foram avaliados comparativamente entre os grupos leigos e ortodontistas, entre os estágios inicial e final do tratamento e entre os grupos tratados com a abordagem cirúrgica e compensatória, com o intuito de estabelecer qual das duas abordagens oferece maior agradabilidade facial. Concluímos que houve proximidade entre as avaliações de leigos e ortodontistas quanto à agradabilidade facial, sendo os leigos mais críticos. Tanto leigos quanto ortodontistas deram escores significativamente maiores para as fotos pós-tratamento nos casos compensatórios e cirúrgicos; os casos cirúrgicos em norma lateral obtiveram os resultados melhores.

Disruption of the CD4–major histocompatibility complex class II interaction blocks the development of CD4+ T cells in vivo

The experiments presented in this report were designed to specifically examine the role of CD4–major histocompatibility complex (MHC) class II interactions during T cell development in vivo. We have generated transgenic mice expressing class II molecules that cannot interact with CD4 but that are otherwise competent to present peptides to the T cell receptor. MHC class II expression was reconstituted in Aβ gene knock-out mice by injection of a transgenic construct encoding either the wild-type I-Aβb protein or a construct encoding a mutation designed to specifically disrupt binding to the CD4 molecule. We demonstrate that the mutation, EA137 and VA142 in the β2 domain of I-Ab, is sufficient to disrupt CD4–MHC class II interactions in vivo. Furthermore, we show that this interaction is critical for the efficient selection of a complete repertoire of mature CD4+ T helper cells as evidenced by drastically reduced numbers of conventional CD4+ T cells in animals expressing the EA137/VA142 mutant I-Ab and by the failure to positively select the transgenic AND T cell receptor on the mutated I-Ab. These results underscore the importance of the CD4–class II interaction in the development of mature peripheral CD4+ T cells.

Cathepsins B and D are dispensable for major histocompatibility complex class II-mediated antigen presentation

Antigen presentation by major histocompatibility complex (MHC) class II molecules requires the participation of different proteases in the endocytic route to degrade endocytosed antigens as well as the MHC class II-associated invariant chain (Ii). Thus far, only the cysteine protease cathepsin (Cat) S appears essential for complete destruction of Ii. The enzymes involved in degradation of the antigens themselves remain to be identified. Degradation of antigens in vitro and experiments using protease inhibitors have suggested that Cat B and Cat D, two major aspartyl and cysteine proteases, respectively, are involved in antigen degradation. We have analyzed the antigen-presenting properties of cells derived from mice deficient in either Cat B or Cat D. Although the absence of these proteases provoked a modest shift in the efficiency of presentation of some antigenic determinants, the overall capacity of Cat B−/− or Cat D−/− antigen-presenting cells was unaffected. Degradation of Ii proceeded normally in Cat B−/− splenocytes, as it did in Cat D−/− cells. We conclude that neither Cat B nor Cat D are essential for MHC class II-mediated antigen presentation.

Differential expression of major histocompatibility complex class II genes on murine macrophages associated with T cell cytokine profile and protective/suppressive effects

Protective/suppressive major histocompatibility complex (MHC) class II alleles have been identified in humans and mice where they exert a disease-protective and immunosuppressive effect. Various modes of action have been proposed, among them differential expression of MHC class II genes in different types of antigen-presenting cells impacting on the T helper type 1 (Th1)–Th2 balance. To test this possibility, the expression of H-2 molecules from the four haplotypes H-2b, H-2d, H-2k, and H-2q was determined on bone marrow-derived macrophages (BMDMs) and splenic B cells. The I-Ab and I-Ek molecules, both well characterized as protective/suppressive, are expressed at a high level on almost all CD11b+ BMDMs for 5–8 days, after which expression slowly declines. In contrast, I-Ad, I-Ak, and I-Aq expression is lower, peaks over a shorter period, and declines more rapidly. No differential expression could be detected on B cells. In addition, the differential MHC class II expression found on macrophages skews the cytokine response of T cells as shown by an in vitro restimulation assay with BMDMs as antigen-presenting cells. The results indicate that macrophages of the protective/suppressive haplotypes express MHC class II molecules at a high level and exert Th1 bias, whereas low-level expression favors a Th2 response. We suggest that the extent of expression of the class II gene gates the back signal from T cells and in this way controls the activity of macrophages. This effect mediated by polymorphic nonexon segments of MHC class II genes may play a role in determining disease susceptibility in humans and mice.

Specific T cell recognition of kinetic isomers in the binding of peptide to class II major histocompatibility complex

Helper T cells are triggered by molecular complexes of antigenic peptides and class II proteins of the major histocompatibility complex . The formation of stable complexes between class II major histocompatibility complex proteins and antigenic peptides is often accompanied by the formation of a short-lived complex. In this report, we describe T cell recognition of two distinct complexes, one short-lived and the other long-lived, formed during the binding of an altered myelin basic protein peptide to I-Ak. One myelin basic protein-specific T cell clone is triggered by only the short-lived complex, and another is triggered by only the stable complex. Thus, a single peptide bound to a particular class II molecule can activate different T cells depending on the conditions of the binding reaction.

Functionality of major histocompatibility complex class II molecules in mice doubly deficient for invariant chain and H-2M complexes

By combining two previously generated null mutations, Ii° and M°, we produced mice lacking the invariant chain and H-2M complexes, both required for normal cell-surface expression of major histocompatibility complex class II molecules loaded with the usual diverse array of peptides. As expected, the maturation and transport of class II molecules, their expression at the cell surface, and their capacity to present antigens were quite similar for cells from Ii°M° double-mutant mice and from animals carrying just the Ii° mutation. More surprising were certain features of the CD4+ T cell repertoire selected in Ii°M° mice: many fewer cells were selected than in Ii+M° animals, and these had been purged of self-reactive specificities, unlike their counterparts in Ii+M° animals. These findings suggest (i) that the peptides carried by class II molecules on stromal cells lacking H-2M complexes may almost all derive from invariant chain and (ii) that H-2M complexes edit the peptide array displayed on thymic stromal cells in the absence of invariant chain, showing that it can edit, in vivo, peptides other than CLIP.

Identification and characterization of a retinoid-induced class II tumor suppressor/growth regulatory gene

Retinoids, synthetic and natural analogs of retinoic acid, exhibit potent growth inhibitory and cell differentiation activities that account for their beneficial effects in treating hyperproliferative diseases such as psoriasis, actinic keratosis, and certain neoplasias. Tazarotene is a synthetic retinoid that is used in the clinic for the treatment of psoriasis. To better understand the mechanism of retinoid action in the treatment of hyperproliferative diseases, we used a long-range differential display–PCR to isolate retinoid-responsive genes from primary human keratinocytes. We have identified a cDNA, tazarotene-induced gene 3 (TIG3; Retinoic Acid Receptor Responder 3) showing significant homology to the class II tumor suppressor gene, H-rev 107. Tazarotene treatment increases TIG3 expression in primary human keratinocytes and in vivo in psoriatic lesions. Increased TIG3 expression is correlated with decreased proliferation. TIG3 is expressed in a number of tissues, and expression is reduced in cancer cell lines and some primary tumors. In breast cancer cell lines, retinoid-dependent TIG3 induction is observed in lines that are growth suppressed by retinoids but not in nonresponsive lines. Transient over-expression of TIG3 in T47D or Chinese hamster ovary cells inhibits colony expansion. Finally, studies in 293 cells expressing TIG3 linked to an inducible promoter demonstrated decreased proliferation with increased TIG3 levels. These studies suggest that TIG3 may be a growth regulator that mediates some of the growth suppressive effects of retinoids.