RESUMEN
Topical 5-aminolevulinic acid-based photodynamic therapy (PDT) has produced complete response rates of >90% for nonmelanoma skin carcinomas, which are mostly human papillomavirus (HPV) negative. Using a similar treatment protocol, we observed a short-term response in only one third (10 of 32) of high-grade vulval intraepithelial neoplasia (VIN 2-3) lesions. Unifocal lesions were found more responsive than multifocal and pigmented lesions. Animal model studies have suggested that long-term PDT response involves an immune reaction in which CTLs play a crucial role. In this study, we have assessed: (a) HPV infection; (b) HLA expression; and (c) immune infiltrating cells in VIN biopsies from responders and nonresponders to determine whether these factors may limit response to topical 5-aminolevulinic acid-based PDT. Tissues from normal vulva (n = 9), vulval carcinoma (n = 11), and VIN (32 patients from which 19 pre- and 43 post-PDT biopsies were taken) were investigated for immune cell infiltration and HLA class I expression by immunohistochemistry and HPV infection by PCR. There was a greater likelihood of HPV positivity associated with a lack of response of VIN to PDT (P = 0.002), and VIN nonresponders were more likely to show HLA class I loss compared with responders (P = 0.030). HLA class I down-regulation was significantly greater in the carcinomas (82%, total loss) than the VIN (28%, 19%, total loss; and 9%, allele loss; P = 0.004). None of the cases with class I down-regulation responded to PDT, whereas 3 of 6 (50%) of cases that showed total class I loss subsequently developed superficial invasion. Compared with normal vulval skin, VIN lesions showed increased infiltration by CD4 (T-helper) and CD68 (macrophages) but not CD1a (Langerhans cells) or CD8 (CTLs). There was, however, a significant increase of CD8 infiltration in posttreatment VIN responders compared with nonresponders (P = 0.0001). These data clearly support the contention that high-risk HPV infection and lack of cell-mediated immunity may play a role in the observed poor response of lower genital lesions to topical PDT.
Asunto(s)
Antígenos HLA/inmunología , Papillomaviridae , Fotoquimioterapia , Neoplasias de la Vulva/inmunología , Neoplasias de la Vulva/virología , Adolescente , Adulto , Anciano , Ácido Aminolevulínico/uso terapéutico , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , ADN Viral/análisis , Femenino , Antígenos HLA/biosíntesis , Antígenos de Histocompatibilidad Clase I/biosíntesis , Antígenos de Histocompatibilidad Clase I/inmunología , Humanos , Células de Langerhans/inmunología , Persona de Mediana Edad , Papillomaviridae/genética , Infecciones por Papillomavirus/complicaciones , Fármacos Fotosensibilizantes/uso terapéutico , Infecciones Tumorales por Virus/complicaciones , Neoplasias de la Vulva/tratamiento farmacológicoAsunto(s)
Antígeno HLA-A2/genética , Mutación , Neoplasias del Cuello Uterino/genética , Neoplasias del Cuello Uterino/inmunología , Empalme Alternativo , Secuencia de Bases , Cartilla de ADN/genética , Exones , Femenino , Expresión Génica , Genes MHC Clase I , Humanos , ARN Mensajero/genética , ARN Mensajero/metabolismo , ARN Neoplásico/genética , ARN Neoplásico/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Neoplasias del Cuello Uterino/metabolismoRESUMEN
Infection with oncogenic human papillomavirus (HPV) types is associated with the development of cervical neoplasia (CIN). The E6 and E7 oncoproteins are constitutively expressed in these lesions and are therefore putative targets for the immune response against HPV. The relation between HPV 16-specific memory cytotoxic T-cell precursor (mCTLp) activity to both oncoproteins and the natural course of cervical dysplasia was analyzed in 38 patients participating in a nonintervention cohort study of women with CIN and 11 HPV 16-positive cervical carcinoma patients. In a cross-sectional study at the end of follow-up prior to biopsy, 8 of 20 patients with a persistent HPV 16 infection had specific mCTLp against at least one of the two oncoproteins. By contrast, no specific mCTLp activity was detected in 11 HPV-negative patients or in 7 patients who had cleared an HPV 16 infection at the end of follow-up. However, 5 of 11 cervical carcinoma patients showed mCTLp activity against the E7 protein only. This study demonstrates that HPV 16 oncogene-specific mCTLp are present in women with HPV 16-positive CIN prior to any intervention. Since HPV-specific mCTLp were detected predominantly in women with high-grade lesions or invasive cervical carcinoma and not in women who cleared the virus, the role of naturally occurring mCTLp in the protection against HPV-associated cervical neoplasia remains to be established.
Asunto(s)
Proteínas Oncogénicas Virales/inmunología , Papillomaviridae/inmunología , Infecciones por Papillomavirus/inmunología , Proteínas Represoras , Linfocitos T Citotóxicos/inmunología , Infecciones Tumorales por Virus/inmunología , Displasia del Cuello del Útero/virología , Neoplasias del Cuello Uterino/virología , Estudios de Cohortes , Estudios Transversales , Femenino , Humanos , Memoria Inmunológica/inmunología , Interleucina-2/biosíntesis , Interleucina-2/metabolismo , Activación de Linfocitos/inmunología , Persona de Mediana Edad , Infecciones por Papillomavirus/virología , Linfocitos T Colaboradores-Inductores/inmunología , Infecciones Tumorales por Virus/virología , Neoplasias del Cuello Uterino/inmunología , Displasia del Cuello del Útero/inmunologíaRESUMEN
The consistent dysregulation of HLA expression in cervical neoplasia is likely to influence the natural history of the disease and prospects for cell-mediated vaccine therapies. We have studied the underlying mechanisms in eight new cervical cancer cell lines derived from primary tumour biopsies. At least five independent mechanisms leading to changes in HLA expression were seen: HLA class I allelic transcription but no protein; abnormal HLA class I allelic transcription; no HLA-B locus transcription; loss of heterozygosity (LOH); no gammaIFN-mediated upregulation of HLA class I expression, and/or no interferon-gamma (gammaIFN)-mediated HLA class II induction. These were evident in different combinations in 7/8 cell lines showing that multiple, mostly irreversible mechanisms not overridden by gammaIFN, are responsible for HLA dysregulation in cervical neoplasia. Point mutations were responsible for lack of HLA-A2 expression in two cases. In cell line 808, the mutation encodes a stop codon in exon 3; in cell line 778, mutation of the first intron acceptor site leads to use of an alternative AG site in exon 2, resulting in a frameshift and a stop codon after the translation of only 38 amino acids. Tumour cells showing specific HLA class I loss may have selective advantage in the face of tumour-specific cytotoxic T cells (CTL). Such immune escape mechanisms present a major obstacle for the success of CTL-mediated therapies in cervical cancer.
Asunto(s)
Carcinoma de Células Escamosas/inmunología , Antígenos de Histocompatibilidad Clase I/inmunología , Proteínas Oncogénicas Virales/inmunología , Neoplasias del Cuello Uterino/inmunología , Transportador de Casetes de Unión a ATP, Subfamilia B, Miembro 2 , Transportadoras de Casetes de Unión a ATP/genética , Transportadoras de Casetes de Unión a ATP/inmunología , Alelos , Biopsia , Carcinoma de Células Escamosas/patología , Carcinoma de Células Escamosas/virología , Cartilla de ADN , Femenino , Regulación Neoplásica de la Expresión Génica/inmunología , Regulación Viral de la Expresión Génica/inmunología , Genotipo , Antígenos de Histocompatibilidad Clase I/genética , Prueba de Histocompatibilidad , Humanos , Proteínas Oncogénicas Virales/genética , Papillomaviridae/genética , Papillomaviridae/inmunología , Infecciones por Papillomavirus/genética , Infecciones por Papillomavirus/inmunología , Fenotipo , Receptores de Interferón/inmunología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN , Linfocitos T Citotóxicos/inmunología , Transcripción Genética/inmunología , Células Tumorales Cultivadas , Infecciones Tumorales por Virus/genética , Infecciones Tumorales por Virus/inmunología , Neoplasias del Cuello Uterino/patología , Neoplasias del Cuello Uterino/virología , Receptor de Interferón gammaRESUMEN
T-helper (Th) cell-dependent IL-2 production and plasma IgG responses to virus-like particles consisting of the human papillomavirus type 16 (HPV-16) major capsid protein L1 (L1-VLP) were determined in patients with cytological evidence of cervical intraepithelial neoplasia (CIN) participating in a non-intervention prospective cohort study. IgG responses were associated with HPV-16 persistence and high-grade CIN lesions, while high frequencies of Th responses were observed in patients with both virus clearance and virus persistence, irrespective of CIN grade. The IgG response was found in conjunction with an IL-2 response to L1-VLP in 87% of the patients. Recognition of the HPV-16 L1 Th epitope (amino acids 311-335) was found to be more closely associated than recognition of L1-VLP as a whole to HPV exposure and CIN development. Among the HPV-16+ patients included in this study, those showing a Th response to amino acids 311-335 were more likely to carry the HLA DRB1*11/DQB1*0301 haplotype, while those showing an IgG response to L1-VLP were more likely to carry DRB1*0101/DQB1*0501. However, neither cell-mediated nor humoral immune responses against HPV-16 L1 appear to be sufficient for the natural control of HPV infection and CIN development.
Asunto(s)
Proteínas de la Cápside , Papillomaviridae/inmunología , Infecciones por Papillomavirus/inmunología , Infecciones Tumorales por Virus/inmunología , Displasia del Cuello del Útero/inmunología , Displasia del Cuello del Útero/virología , Neoplasias del Cuello Uterino/inmunología , Neoplasias del Cuello Uterino/virología , Adulto , Secuencia de Aminoácidos , Anticuerpos Antivirales/sangre , Antígenos Virales/genética , Estudios de Cohortes , Epítopos/genética , Femenino , Genotipo , Antígenos HLA-DQ/genética , Cadenas beta de HLA-DQ , Antígenos HLA-DR/genética , Cadenas HLA-DRB1 , Haplotipos , Humanos , Inmunoglobulina G/sangre , Interleucina-2/biosíntesis , Datos de Secuencia Molecular , Proteínas Oncogénicas Virales/genética , Proteínas Oncogénicas Virales/inmunología , Papillomaviridae/clasificación , Papillomaviridae/genética , Infecciones por Papillomavirus/complicaciones , Linfocitos T Colaboradores-Inductores/inmunología , Infecciones Tumorales por Virus/complicaciones , Neoplasias del Cuello Uterino/complicaciones , Displasia del Cuello del Útero/complicacionesRESUMEN
The considerable morbidity and mortality associated with certain human papillomaviruses (HPV) has provided the impetus for HPV vaccine development. The design of such vaccines has evolved from an understanding of the nature of HPV infections and their consequences, together with evaluation of the efficacy of different approaches to vaccination in animal models. These studies have culminated in the production of several different vaccine preparations which are currently undergoing Phase I and II clinical trials. The justification for the widespread implementation of prophylactic HPV vaccines will depend on the outcome of larger scale studies of vaccine efficacy that take into account the epidemiology of HPV infections and associated disease. The usefulness of therapeutic HPV vaccines will require evidence that they can substantially augment or substitute for the effectiveness of currently available treatments.
Asunto(s)
Antígenos Virales/inmunología , Papillomaviridae/inmunología , Infecciones por Papillomavirus/prevención & control , Vacunas contra Papillomavirus , Infecciones Tumorales por Virus/prevención & control , Vacunas Virales , Bacterias/genética , Ensayos Clínicos como Asunto , ADN Viral/inmunología , Células Dendríticas/inmunología , Modelos Animales de Enfermedad , Femenino , Humanos , Inmunidad Innata , Papillomaviridae/genética , Papillomaviridae/crecimiento & desarrollo , Papillomaviridae/patogenicidad , Infecciones por Papillomavirus/terapia , Péptidos/inmunología , Infecciones Tumorales por Virus/terapia , Neoplasias del Cuello Uterino/prevención & control , Neoplasias del Cuello Uterino/terapia , Neoplasias del Cuello Uterino/virología , Vacunas de ADN/inmunología , Proteínas Virales/inmunología , Vacunas Virales/economía , Virión/inmunologíaRESUMEN
High-risk human papillomavirus (HPV) infection plays an important role in cervical intra-epithelial neoplasia (CIN), but HPV infection alone is not sufficient for progression to cervical cancer. Several lines of evidence suggest that cellular immune surveillance is important in the control of HPV infection and the development of CIN. The presentation to T cells of target viral peptides in the context of HLA molecules is influenced by the genetic polymorphisms of both HPV and HLA and thereby influences the host immune response and clinical outcome of HPV infection. HLA class I and II polymorphism in susceptibility for HPV 16 infection, development and progression of CIN was analyzed in a group of 118 patients participating in a prospective study of women with initial abnormal cytology. Patients were stratified according to HPV status and course of the disease. HLA-B*44 frequency was increased in the small group of patients with a lesion that showed clinical progression during follow-up [OR = 9.0 (4.6-17.5), p = 0.007]. HLA-DRB1*07 frequency was increased among HPV 16-positive patients compared with patients who were negative for all HPV types [OR = 5.9 (3.0-11.3), p = 0.02]. Our results are consistent with the immunogenetic factors associated with disease progression being different from those associated with susceptibility to HPV 16 infection. Sequencing of the HPV 16 E6 and E7 open reading frames of a subset of these patients (n = 40) showed the frequency of HPV 16 variants to be similar to other studies. However, there was no significant correlation between variant incidence and disease progression or viral persistence and no significant correlation with any HLA allele. It appears that multiple HLA types can influence HPV 16-associated cervical dysplasia but the role of HPV 16 variants in disease progression and susceptibility in relation to HLA polymorphism remains unclear.
Asunto(s)
Antígenos de Histocompatibilidad Clase II/genética , Antígenos de Histocompatibilidad Clase I/genética , Proteínas Oncogénicas Virales/genética , Papillomaviridae/genética , Infecciones por Papillomavirus/complicaciones , Proteínas Represoras , Infecciones Tumorales por Virus/complicaciones , Displasia del Cuello del Útero/virología , Neoplasias del Cuello Uterino/virología , Adulto , Alelos , Progresión de la Enfermedad , Femenino , Variación Genética , Genotipo , Humanos , Persona de Mediana Edad , Sistemas de Lectura Abierta , Proteínas E7 de Papillomavirus , Infecciones por Papillomavirus/genética , Infecciones por Papillomavirus/virología , Polimorfismo Genético , Estudios Prospectivos , Infecciones Tumorales por Virus/genética , Infecciones Tumorales por Virus/virología , Neoplasias del Cuello Uterino/patología , Displasia del Cuello del Útero/patologíaRESUMEN
T-cell-mediated immune responses against oncogenic human papillomaviruses (HPVs) are believed to play a role in the prevention of cervical carcinogenesis. The in vitro production of interleukin 2 by CD4+ T helper (Th) cells in response to overlapping 20-mer peptides covering the HPV-16 E7 oncoprotein sequence was determined in 72 women with cytological evidence of premalignant cervical intraepithelial neoplasia (CIN) who participated in a nonintervention follow-up (FU) study. In addition, 15 HPV-16 + cervical carcinoma patients were tested. Positive Th cell reactivity was restricted to patients infected by HPV-16 and related types and showed a strong association with viral persistence and disease progression, as evidenced by the high frequency of positive responders among women with persistent HPV-16 infections who ended FU with high-grade CIN III lesions [14 of 15 (93%)]. Women with cervical carcinoma showed responses at a significantly reduced rate [7 of 15 (47%); P = 0.014]. Over the FU period (10-34 months), the level of E7-induced interleukin 2 production from the lymphocytes of CIN patients who had cleared HPV-16 infection showed an inverse correlation with time relative to the last positive HPV DNA test, with 8 of 13 of these patients showing positive responses after clearance. By contrast, among women with persistent HPV-16 infections and developing CIN III lesions (n = 8), there was a rise in Th cell activity over the course of FU. The majority of women responded to an immunogenic region in the carboxyl terminus of the E7 protein (amino acids 67-98). The observed HPV-16 E7-specific Th cell responses may develop as a consequence of increased antigen availability resulting either from clearance or from progression of cervical lesions.
Asunto(s)
Carcinoma/metabolismo , Interleucina-2/metabolismo , Proteínas Oncogénicas Virales/farmacología , Linfocitos T Colaboradores-Inductores/metabolismo , Displasia del Cuello del Útero/metabolismo , Neoplasias del Cuello Uterino/metabolismo , Adulto , Carcinoma/genética , Carcinoma/patología , Carcinoma/virología , Células Cultivadas , Femenino , Antígenos HLA-DQ/genética , Antígenos HLA-DR/genética , Humanos , Leucocitos Mononucleares/metabolismo , Estudios Longitudinales , Persona de Mediana Edad , Proteínas E7 de Papillomavirus , Estudios Prospectivos , Linfocitos T Colaboradores-Inductores/virología , Neoplasias del Cuello Uterino/genética , Neoplasias del Cuello Uterino/patología , Neoplasias del Cuello Uterino/virología , Displasia del Cuello del Útero/genética , Displasia del Cuello del Útero/patología , Displasia del Cuello del Útero/virologíaRESUMEN
HLA class I downregulation is a frequent event associated with tumour invasion and development. Altered HLA class I tumour phenotypes can have profound effects on T-cell and natural killer (NK)-cell antitumour responses. Here, Federico Garrido and colleagues analyse these altered tumour phenotypes in detail, indicating their potential relevance for implementation of immunotherapeutic protocols and strategies to overcome tumour escape mechanisms.
Asunto(s)
Antígenos HLA/clasificación , Antígenos de Histocompatibilidad Clase I/clasificación , Vigilancia Inmunológica , Inmunofenotipificación , Neoplasias/inmunología , HumanosRESUMEN
No HLA allele or specificity was significantly different in frequency between a group of 150 cervical cancer patients from north-west England and controls (corrected P values). HLA-DRB1*1501/DQB1*0602 was non-significantly increased, particularly among patients with HPV16-positive tumours. HLA-B7-positive patients had a significantly poorer clinical outcome than HLA-B7-negative patients. A significant component of the genotypic effect is down-regulation of HLA-B7 expression by the tumour cells.
Asunto(s)
Prueba de Histocompatibilidad , Papillomaviridae/clasificación , Infecciones por Papillomavirus/inmunología , Infecciones Tumorales por Virus/inmunología , Neoplasias del Cuello Uterino/inmunología , Adulto , Anciano , Anciano de 80 o más Años , Progresión de la Enfermedad , Femenino , Estudios de Seguimiento , Humanos , Inmunogenética , Persona de Mediana Edad , Papillomaviridae/genética , Infecciones por Papillomavirus/virología , Infecciones Tumorales por Virus/virología , Neoplasias del Cuello Uterino/virologíaRESUMEN
The pattern of cell surface antigen expression of a set of cell lines derived from human germ cell tumours and corresponding to various cell phenotypes found within these tumours was studied using immunofluorescence. Twenty-two different antibodies were used. Many of these antibodies have been noted to recognise epitopes that are either preferentially expressed by embryonal carcinoma (EC) cells, or by more differentiated cell types. Using scatter plots and rank correlations, 6 groups of antibodies were distinguished with respect to their staining patterns on the cell lines tested. Several antibodies showed a specific staining pattern in relation to the differentiation state of the cells. Two groups of antibodies included those recognising high m.w. glycoproteins (antibodies TRA-1-60, TRA-1-81, GCTM2, 3-177, K4 and K21) and the ganglioseries glycolipid antigens SSEA-3 and -4 (antibodies MC631 and MC813-70). These antibodies mostly stained EC cells but not other cell types, confirming previously published data. However, one of these groups, comprising antibodies K4 and MC631, was more exclusively associated with the EC cell phenotype than was the other group. Antibodies recognising the liver isozyme of alkaline phosphatase (TRA-2-49 and TRA-2-54) also reacted strongly with most EC cell lines, although they reacted significantly with a number of other cell lines as well, whereas antibodies to the placental isozyme tended to react only weakly with EC cells. The antibodies recognising the ganglioseries glycolipids GD2 and GD3 (VIN2PB22 and VINIS56) preferentially stained cells with neuroectodermal characteristics. Other antibodies showed a heterogeneous staining pattern for the cell lines with different phenotypes. The data obtained from the cell lines were, in general, similar to data obtained from immunohistochemical studies on tissue sections of primary germ cell tumours of the adult testis, including carcinoma in situ.
Asunto(s)
Antígenos de Neoplasias/análisis , Antígenos de Superficie/análisis , Germinoma/inmunología , Neoplasias Testiculares/inmunología , Adulto , Fosfatasa Alcalina/inmunología , Anticuerpos Monoclonales/clasificación , Anticuerpos Monoclonales/inmunología , Anticuerpos Antineoplásicos/clasificación , Anticuerpos Antineoplásicos/inmunología , Especificidad de Anticuerpos , Biomarcadores de Tumor/inmunología , Biopsia , Carcinoma in Situ/química , Carcinoma in Situ/inmunología , Carcinoma in Situ/patología , Carcinoma Embrionario/química , Carcinoma Embrionario/inmunología , Carcinoma Embrionario/patología , Tumor del Seno Endodérmico/química , Tumor del Seno Endodérmico/inmunología , Tumor del Seno Endodérmico/patología , Citometría de Flujo , Técnica del Anticuerpo Fluorescente Indirecta , Expresión Génica , Germinoma/química , Germinoma/clasificación , Germinoma/patología , Glucolípidos/inmunología , Humanos , Inmunofenotipificación , Isoenzimas/inmunología , Masculino , Glicoproteínas de Membrana/inmunología , Microscopía Fluorescente , Proteínas de Neoplasias/inmunología , Seminoma/química , Seminoma/inmunología , Seminoma/patología , Neoplasias Testiculares/química , Neoplasias Testiculares/clasificación , Neoplasias Testiculares/patología , Testículo/patología , Células Tumorales CultivadasRESUMEN
The development of cervical carcinoma is strongly associated with specific types of human papillomaviruses (HPVs). A role for cellular immunity in cervical disease is supported by the increased occurrence of HPV-associated lesions in immunosuppressed individuals. Upon viral infection or malignant transformation, ensuing alterations in gene expression result in the generation of novel sets of peptides which can form complexes with specific HLA class I heavy chains and beta 2-microglobulin. These are then expressed at the cell surface as potential targets for specific T cells. In this study of 100 carcinomas HLA-A and -B class I expression by the tumour cells was down-regulated at one or more alleles in at least 73% of cervical carcinomas. Interference with the transporter associated with antigen presentation (TAP), which translocates cytosolic peptides from endogenously synthesised proteins (e.g. viral) into the lumen of the endoplasmic reticulum was found in 38% of the HLA class I down-regulated tumours. Loss of expression for common HLA class I alleles ranged from 36% to 71%, and such changes might be expected to influence specific immunogenic peptide presentation and consequent immune recognition. These results underline the importance of single as well as multiple allelic loss in cervical neoplasia and have important implications for attempts to intervene immunologically in cervical cancer.
Asunto(s)
Transportadoras de Casetes de Unión a ATP/genética , Alelos , Regulación hacia Abajo , Regulación Neoplásica de la Expresión Génica , Antígenos HLA-A/genética , Antígenos HLA-B/genética , Neoplasias del Cuello Uterino/genética , Transportador de Casetes de Unión a ATP, Subfamilia B, Miembro 2 , Adenocarcinoma/genética , Adulto , Anciano , Anciano de 80 o más Años , Biopsia , Carcinoma de Células Escamosas/genética , Cuello del Útero/fisiología , Femenino , Eliminación de Gen , Heterocigoto , Humanos , Persona de Mediana Edad , Valores de Referencia , Neoplasias del Cuello Uterino/inmunologíaRESUMEN
HLA-restricted cytotoxic T-lymphocyte (CTL) recognition of human papillomavirus (HPV) oncogene products may be important in the control of the HPV infections associated with the development of cervical cancer. We have identified, in HLA-B7 individuals, a consistent variation in the HPV16 E6 oncoprotein sequence, which alters an HLA-B7 peptide binding epitope in a way likely to influence immune recognition by CTLs. These results illustrate a biologically relevant mechanism for escape from immune surveillance of HPV16 in HLA-B7 individuals. Thus, both HLA type and HPV16 strain variation need to be considered in the screening of at-risk individuals and for the rational design of anti-HPV vaccines.
Asunto(s)
Antígeno HLA-B7/inmunología , Proteínas Oncogénicas Virales/inmunología , Papillomaviridae/inmunología , Proteínas Represoras , Neoplasias del Cuello Uterino/inmunología , Vacunas Virales/inmunología , Secuencia de Aminoácidos , Epítopos/análisis , Femenino , Humanos , Datos de Secuencia Molecular , Mutación/inmunología , Unión Proteica/inmunología , Análisis de Secuencia de ADN , Linfocitos T Citotóxicos/inmunología , Factores de Transcripción/inmunología , Neoplasias del Cuello Uterino/terapiaRESUMEN
An association of HLA-DQ3 with SCC of the cervix has been reported by researchers in Germany and Norway. This article documents a similar-sized study with patients and controls from northwest England. We report in detail on serologically determined HLA polymorphism in SCC patients with respect to HPV 16 infection, MHC class II expression within the tumor, serologic response to HPV, and other relevant clinical variables. We have also extended our studies to include DNA-based analysis using PCR and SSO probes for HLA-DQ. No significant association of any HLA-A, -B, -C, -DR, or -DQ antigen with SCC patients was found. While a possible explanation of the differences among studies could be a reflection of disease heterogeneity, the several tumor and clinical factors examined do not account for the observed differences from previous reports. Further studies are needed for a greater understanding of the interaction of HPV and HLA type in the development of cervical neoplasia.
Asunto(s)
Carcinoma de Células Escamosas/inmunología , Antígenos HLA/análisis , Papillomaviridae , Infecciones por Papillomavirus/inmunología , Infecciones Tumorales por Virus/inmunología , Neoplasias del Cuello Uterino/inmunología , Adulto , Anciano , Anciano de 80 o más Años , Alelos , Carcinoma de Células Escamosas/microbiología , ADN Viral/análisis , Femenino , Antígenos HLA/genética , Antígenos HLA-DQ/análisis , Antígenos HLA-DQ/genética , Humanos , Persona de Mediana Edad , Papillomaviridae/genética , Infecciones por Papillomavirus/microbiología , Reacción en Cadena de la Polimerasa , Polimorfismo Genético , Infecciones Tumorales por Virus/microbiología , Neoplasias del Cuello Uterino/microbiologíaRESUMEN
Plasma cells within bone marrow aspirates from multiple myeloma patients have been shown to be reactive with the lectin peanut agglutinin (PNA). This has been recently exploited by using PNA for purging bone marrow of malignant cells in autotransplantation therapy of the disease. The purpose of this investigation was to isolate and characterize the PNA-binding proteins of myeloma cells. We used the malignant plasma cell-derived line Karpas-620 (K620) as a model, and showed by affinity chromatography, SDS-PAGE, and immunoprecipitation that, among several PNA-binding proteins, a major one is an incompletely sialylated form of CD44. CD44 is a well-known homing receptor protein which is rich in carbohydrate and usually completely sialylated so that it does not react with PNA. We have then examined the PNA reactivity of myeloma cells from different patients and showed a clear difference in the profile of PNA-binding proteins from case to case. Moreover, in contrast to K620 cells, some of the patient plasma cells tested did not have a PNA-binding form of CD44. In conclusion, therefore, we have shown that a number of different proteins participate in PNA binding by malignant plasma cells. Moreover, we have demonstrated a novel, incompletely sialylated form of CD44 on a myeloma cell line. It is known that the level of glycosylation of CD44 and other proteins may affect their function, but how this relates to the malignant behaviour of plasma cells remains to be determined.
Asunto(s)
Lectinas/metabolismo , Mieloma Múltiple/metabolismo , Células Plasmáticas/metabolismo , Médula Ósea/metabolismo , Purgación de la Médula Ósea , Cromatografía de Afinidad , Electroforesis en Gel de Poliacrilamida , Humanos , Aglutinina de Mani , Receptores Mensajeros de Linfocitos/metabolismo , Células Tumorales CultivadasRESUMEN
The observation that tumor cells of some neoplasms display major histocompatibility complex (MHC) class II molecules may be of functional significance, influencing the progression of malignancy by allowing the cancer cells to present antigen to the immune system. In the normal cervix, class II molecules are expressed by columnar but not squamous epithelium. The pattern of MHC class II expression in cervical carcinomas has been documented using immunohistochemical methods. Of 53 cervical squamous carcinomas examined for MHC class II expression, only 17% maintained a negative phenotype characteristic of the epithelium from which they were derived, while the remaining tumors exhibited either uniform (45%) or heterogeneous (38%) expression. Tumor areas which were class II positive also express class II associated invariant chain and the adhesion molecules lymphocyte function antigen 3 and intercellular adhesion molecule 1. The DR, DP, and DQ class II MHC subloci are differentially expressed, suggesting independent regulation. There is a trend for tumors with the uniform class II phenotype to predominantly express DR antigen, whereas tumors of the heterogeneous class II phenotype express with equal frequency either DR or DP antigens dominantly. There is no apparent influence of class II status on lymphocyte infiltration of the tumors. The presence of human papillomavirus 16 DNA in the cervical carcinoma specimens was analyzed by Southern blotting of restriction enzyme digested DNA and no correlation between the presence of human papilloma virus and MHC class II expression was found.
Asunto(s)
Antígenos de Histocompatibilidad Clase II/análisis , Papillomaviridae , Infecciones Tumorales por Virus/inmunología , Neoplasias del Cuello Uterino/inmunología , Adulto , Anciano , Antígenos de Superficie/análisis , Antígenos CD58 , Carcinoma de Células Escamosas/inmunología , Moléculas de Adhesión Celular/análisis , Cuello del Útero/inmunología , ADN Viral/análisis , Femenino , Antígenos HLA-DP/análisis , Antígenos HLA-DQ/análisis , Antígenos HLA-DR/análisis , Antígenos de Histocompatibilidad Clase I/análisis , Humanos , Molécula 1 de Adhesión Intercelular , Recuento de Leucocitos , Linfocitos Infiltrantes de Tumor/inmunología , Glicoproteínas de Membrana/análisis , Persona de Mediana Edad , Papillomaviridae/genética , Fenotipo , Infecciones Tumorales por Virus/complicaciones , Neoplasias del Cuello Uterino/etiologíaRESUMEN
A significant decrease in the frequency of BF*F allele and an increase of BF*F1 allele was found in 101 clinically definite multiple sclerosis patients compared to 270 normal controls from North-East England. In Dw2 types 41 patients and 60 controls, only the rare allele BF*F1 showed a significant increase in the patients group. For the common BF*S allele a significant increase was found in Dw2+ patients compared to the Dw2- patients, but a slight similar increase observed in Dw2+ controls did not attain significance. This increase in the patient group is attributed to a strong linkage disequilibrium between BF*S and Dw2 alleles. No such linkage disequilibrium exists in the normal controls. There is a suggestion that the BF*S and Dw2+ alleles are more prevalent in chronic progressive patients, implying that in Dw2+ patients BF may influence the progression of the disease.
Asunto(s)
Alelos , Factor B del Complemento/genética , Antígenos HLA-D/genética , Esclerosis Múltiple/genética , Inglaterra , Frecuencia de los Genes , Humanos , FenotipoRESUMEN
The IgG subclasses secreted by human B cells in vitro in response to IL-2 have been analysed. B cells were prepared from tonsil, blood and spleen, and cultured with recombinant IL-2 in the presence or absence of two polyclonal activators: Staphylococcus aureus Cowan 1 (SAC) and bacterial lipopolysaccharide (LPS). Secretion of all four subclasses and of IgM was stimulated by IL-2, but the relative amounts varied according to (i) the tissue source of the B cells, and (ii) which polyclonal activator was used. The amount of IgG1 tended to be higher and IgG2 tended to be lower when SAC was the polyclonal activator (compared to LPS). This difference was most marked for tonsil B cells, and it was found that SAC had a negative effect on secretion of IgM and IgG2 in these cultures, whilst synergizing with IL-2 to stimulate the production of IgG1, 3 and 4. When the degree of stimulation of different pairs of isotypes was analysed, several interesting positive correlations emerged. In tonsil B-cell cultures, stimulation of IgM and IgG2 was linked with each other, but not with IgG1, whilst in blood B-cell cultures all isotypes appeared to be stimulated co-ordinately. Stimulation of IgG1 and IgG3 were positively correlated in cultures of B cells from all tissues. The results emphasize that the effects of a single cytokine on immunoglobulin isotype production can be influenced by the source of the B cells, and by other signals delivered to the cells.
Asunto(s)
Antígenos Bacterianos/inmunología , Linfocitos B/inmunología , Inmunoglobulina G/inmunología , Interleucina-2/inmunología , Células Cultivadas , Humanos , Inmunoglobulina G/biosíntesis , Interleucina-2/farmacología , Lipopolisacáridos/inmunología , Activación de Linfocitos/inmunología , Tonsila Palatina/inmunología , Staphylococcus aureus/inmunologíaRESUMEN
Factor VIII concentrate inhibits T-cell function in vitro and in vivo. The mechanisms underlying the phenomenon were investigated. Factor VIII concentrate has a direct effect on lymphocytes, uninfluenced by haemophilic monocyte dysfunction, since it inhibited lymphocyte transformation with phorbol myristate acetate, a reaction unaffected by monocyte depletion. Inhibition of lymphocyte transformation by factor VIII concentrate is not corrected by the addition of exogenous IL2, suggesting that it does not inhibit lymphocyte function by suppression of IL2 secretion alone. Factor VIII concentrate causes profound inhibition of IL2-receptor expression (CD25); with an 89% reduction in CD25-positive CD4 cells and a 50% reduction in CD25-antigen molecules per cell. CD8 lymphocytes are similarly affected. Smaller reductions in CD71 and HLA-DR expression are also observed. Down modulation of CD25-antigen may explain the reduced IL2 secretion observed by others, and may be an important cause of immunodeficiency in HIV-seronegative haemophiliacs.
Asunto(s)
Factor VIII/inmunología , Hemofilia A/inmunología , Tolerancia Inmunológica , Receptores de Interleucina-2/análisis , Antígenos CD/análisis , Relación Dosis-Respuesta Inmunológica , Antígenos HLA-DR/análisis , Humanos , Interleucina-2/inmunología , Activación de Linfocitos , Monocitos/inmunologíaRESUMEN
Lymphocyte function and cell surface phenotype were examined in fifteen patients with late onset hypogammaglobulinaemia. The percentage of surface immunoglobulin-positive B cells in fourteen of the fifteen patients was in the normal range. Patients' B cells expressed MHC class II antigens at normal levels. For one patient, there was relatively high sIgD and low sIgM expression on B cells; the rest of the patients did not differ from controls in surface immunoglobulin density. The proportion of B cells positive for CD5 in patients was comparable to normal controls, and considerably less than in cord blood. However, the pattern of immunoglobulin isotype secretion in vitro by patients' B cells closely paralleled responses of cord blood B cells. Spontaneous secretion of IgM and IgG by patients' B cells was very low. Following polyclonal activation in the presence of autologous T cells, cells from thirteen patients secreted IgM within the normal range in response to at least one activator. The response of patients' purified B cells to IL-2 and gamma-IFN was variable. For four of six tested, B cells cultured with IL-2 and gamma-IFN together with polyclonal activators secreted normal levels of IgM. B cells from the other two patients secreted little or no IgM in response to these cytokines. For fourteen patients, IgG secretion following polyclonal activation remained low both when B cells were cultured with T cells or with a combination of IL-2 and gamma-IFN. IgG subclass imbalance was seen in one patient, whose cells secreted an unusually high proportion of IgG3, and undetectable IgG2 and IgG4; this pattern was consistent whether T cell help was provided by autologous or allogeneic T cells. Similarly purified B cells from this patient showed deficient IgG2 and IgG4 production in response to IL-2 and gamma-IFN.