RESUMO
PURPOSE: To evaluate the feasibility and sensitivity of multimodality PET/CT and MRI imaging for non-invasive characterization of brain microglial/macrophage activation occurring during the acute phase in a mouse model of relapsing remitting multiple sclerosis (RR-MS) using [18F]DPA-714, a selective radioligand for the 18-kDa translocator protein (TSPO), superparamagnetic iron oxide particles (SPIO), and ex vivo immunohistochemistry. METHODS: Experimental autoimmune encephalomyelitis (EAE) was induced in female SJL/J mice by immunization with PLP139-151. Seven symptomatic EAE mice and five controls underwent both PET/CT and MRI studies between 11 and 14 days post-immunization. SPIO was injected i.v. in the same animals immediately after [18F]DPA-714 and MRI acquisition was performed after 24 h. Regional brain volumes were defined according to a mouse brain atlas on co-registered PET and SPIO-MRI images. [18F]DPA-714 standardized uptake value (SUV) ratios (SUVR), with unaffected neocortex as reference, and SPIO fractional volumes (SPIO-Vol) were generated. Both SUVR and SPIO-Vol values were correlated with the clinical score (CS) and among them. Five EAE and four control mice underwent immunohistochemical analysis with the aim of identifying activated microglia/macrophage and TSPO expressions. RESULTS: SUVR and SPIO-Vol values were significantly increased in EAE compared with controls in the hippocampus (p < 0.01; p < 0.02, respectively), thalamus (p < 0.02; p < 0.05, respectively), and cerebellum and brainstem (p < 0.02), while only SPIO-Vol was significantly increased in the caudate/putamen (p < 0.05). Both SUVR and SPIO-Vol values were positively significantly correlated with CS and among them in the same regions. TSPO/Iba1 and F4/80/Prussian blue staining immunohistochemistry suggests that increased activated microglia/macrophages underlay TSPO expression and SPIO uptake in symptomatic EAE mice. CONCLUSIONS: These preliminary results suggest that both activated microglia and infiltrated macrophages are present in vulnerable brain regions during the acute phase of PLP-EAE and contribute to disease severity. Both [18F]DPA-714-PET and SPIO-MRI appear suitable modalities for preclinical study of neuroinflammation in MS mice models.
Assuntos
Encefalomielite Autoimune Experimental , Animais , Encefalomielite Autoimune Experimental/diagnóstico por imagem , Feminino , Ativação de Macrófagos , Macrófagos , Imageamento por Ressonância Magnética , Camundongos , Microglia , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada , Tomografia por Emissão de Pósitrons , Pirazóis , PirimidinasRESUMO
Cell-mediated immune response to breast tumor has only been marginally investigated. To gain insight into this issue we have developed two clones of distinct phenotype, CD3+ alpha/beta, CD4+, CD8-, CD16-, and CD3+ alpha/beta, CD4-, CD8+, CD16-, respectively, from peripheral blood lymphocytes (PBL) of a breast cancer patient. These effectors, selected on the basis of their cytolytic activity against autologous tumor cells and lack of lysis on NK-sensitive cell lines, preferentially recognize autologous tumor cells. The two clones' cytotoxic activity, while inhibited by anti-LFA-1 mAb, could not be abolished by mAbs to CD3, to class I and class II MHC molecules, and by mAbs to molecules involved in T cell function (i.e., CD4, CD8, CD2). The molecular structure of the alpha and beta T cell receptor chains of the two effector cells, confirmed their clonality and showed that, despite an overlapping killing pattern, they possess distinct TCR alpha and beta chains. These findings demonstrate that breast tumor-specific CTL clones can be generated through current technology and that a alpha/beta effector cell population operating through a HLA-unrestricted and TCR/CD3-independent pathway may be involved in the identification and killing of this tumor.
Assuntos
Neoplasias da Mama/imunologia , Receptores de Antígenos de Linfócitos T alfa-beta/imunologia , Subpopulações de Linfócitos T/imunologia , Linfócitos T Citotóxicos/imunologia , Sequência de Aminoácidos , Antígenos de Diferenciação de Linfócitos T/imunologia , Sequência de Bases , Feminino , Antígenos de Histocompatibilidade Classe I/imunologia , Antígenos de Histocompatibilidade Classe II/imunologia , Humanos , Imunidade Celular , Teste de Cultura Mista de Linfócitos , Pessoa de Meia-Idade , Dados de Sequência Molecular , RNA Mensageiro/análise , Receptores de Antígenos de Linfócitos T alfa-beta/genética , Alinhamento de Sequência , Análise de Sequência de DNA , Células Tumorais CultivadasRESUMO
Although much effort has been expended on evaluating recombinant proteins and synthetic peptides as immunogens, they have generally proved incapable of inducing an efficient cytotoxic T-cell (CTL) response. Filamentous bacteriophage fd can display multiple copies of foreign peptides in the N-terminal region of its major coat protein pVIII, 2,700 copies of which make up the virus capsid. Here we show that fd virions displaying peptide RT2 (ILKEPVHGV), corresponding to residues 309-317 of the reverse transcriptase (RTase) of HIV-1, are able to prime a CTL response specific for this HIV-1 epitope in human cell lines. Successful priming also requires a T-helper epitope, pep23 (KDSWTVNDIQKLVGK), corresponding to residues 249-263 of HIV-1 RTase. Supplying this by displaying it on either the same or a separate bacteriophage virion led to activation of antigen-specific CD4+ T cells. Likewise, HLA-A2 transgenic mice immunized with bacteriophage virions displaying peptide RT2 were shown to mount an effective, specific anti-HIV-RT2 CTL response. This unexpected ability to elicit a designated cytolytic T-cell response, in addition to a B-cell response, has important implications for access to the class I major histocompatibility complex (MHC) loading compartment and the development of recombinant vaccines.
Assuntos
Epitopos/genética , HIV-1/imunologia , Inovirus/genética , Sequência de Aminoácidos , Animais , Células Cultivadas , Humanos , Camundongos , Subpopulações de Linfócitos T , Linfócitos T Citotóxicos/citologia , Linfócitos T Citotóxicos/imunologiaRESUMO
A systematic exploration of the V(H)2/V(kappa)12-13 variable domains of the anti-CD4 monoclonal antibody (mAb) 13B8.2 was performed by the Spot method to screen for paratope-derived peptides (PDPs) demonstrating CD4 binding ability. Nine peptides, named CB1 to CB9, were identified, synthesized in a cyclic and soluble form and tested for binding to recombinant soluble CD4. Among them, CB1, CB2 and CB8 showed high anti-CD4 activity. Competition studies for CD4 binding indicated that PDPs CB1, CB8, and the parental mAb 13B8.2 recognized the same complementarity determining region (CDR)3-like loop region. PDP CB1 was shown to mimic the biological properties of 13B8.2 mAb in two independent cellular assays, demonstrating inhibitory activities in the micromolar range on antigen presentation and human immunodeficiency virus promoter activation. Our results indicate that the bioactive CDR-H1 PDP CB1 has retained a significant part of the parental 13B8.2 mAb properties and might be a lead for the design of anti-CD4 peptidomimetics of clinical interest.
Assuntos
Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/metabolismo , Antígenos CD4/imunologia , Fragmentos de Peptídeos/imunologia , Animais , Fármacos Anti-HIV/imunologia , Fármacos Anti-HIV/metabolismo , Fármacos Anti-HIV/farmacologia , Anticorpos Monoclonais/farmacologia , Apresentação de Antígeno , Antígenos CD4/metabolismo , Linhagem Celular , Ensaio de Imunoadsorção Enzimática , Epitopos , Regulação da Expressão Gênica , Genes Reporter , HIV-1/metabolismo , Humanos , Região Variável de Imunoglobulina , Imunossupressores/metabolismo , Imunossupressores/farmacologia , Interleucina-2/metabolismo , Camundongos , Mimetismo Molecular , Dados de Sequência Molecular , Fragmentos de Peptídeos/metabolismo , Regiões Promotoras Genéticas , Ligação Proteica , Proteínas Recombinantes de Fusão/metabolismoRESUMO
T cell clones are an irreplaceable asset for the study of immune responses relevant to human pathologies. Such cells, however, cannot always be maintained in long-term culture. In order to reconstitute functional human T cell receptors (TCRs) into stable and fast growing hybridoma T cells, we developed a general approach based on a versatile cassette system, which allows cloning of all types of human T cell receptor variable alpha and beta region genes fused to murine constant regions. These chimeric constructs are easily excised and transferred into expression vectors that can be used to transfect a human CD4-expressing murine T cell hybridoma recipient. The resulting transfectants are highly stable both in terms of T cell receptor-CD3 expression and IL-2 response to the specific antigenic stimulus. Using these cassette vectors, we reconstituted the original HLA-restricted antigen specificity for two human T cell clones, one recognizing an immunodominant epitope of HIV-1 gp120, and the other recognizing an immunodominant epitope of HIV-1 reverse transcriptase. We found that the reconstituted hybridomas maintain the ability of the original T cell clones to recognize the appropriate epitope in the context of the relevant MHC either as a synthetic peptide or after processing. Their unlimited growth capacity makes them particularly suited for in vitro studies.
Assuntos
Clonagem Molecular/métodos , Vetores Genéticos/genética , Fragmentos de Peptídeos/genética , Receptores de Antígenos de Linfócitos T alfa-beta/genética , Animais , Proteína gp120 do Envelope de HIV/imunologia , Transcriptase Reversa do HIV/imunologia , Humanos , Hibridomas , Epitopos Imunodominantes/imunologia , Camundongos , Mutagênese Insercional , Proteínas Recombinantes de FusãoRESUMO
In recent years, a growing interest in the study of peptide antigenicity in relation to the role of flanking sequences and protein topology in processing, presentation, and recognition has been observed. However, the information available on the antigenicity of recombinant fusion proteins and their effect on the selection of antigen receptor repertoires is limited. To analyze the role of molecular topology of T epitopes in a system relevant to human pathology, we have used the bacterially expressed Schistosoma japonicum glutathione S transferase (GST) to construct recombinant antigens containing HIV-1 derived T cell determinants, and human T cell clones specific for these determinants. We found that antigenicity of a given GST-peptide combination was not the same when T cells and antigen presenting cells from different individuals were tested. Our results show that differences in processing and presentation of chimeric proteins are not dictated by the use of diverse restriction elements. We also found that the context in which an antigenic peptide is delivered affects the recruited repertoire as defined according to T cell receptor V beta usage and fine specificities of selected T cells.
Assuntos
Apresentação de Antígeno , Epitopos/genética , Epitopos/imunologia , Linfócitos T/imunologia , Antígenos Virais/genética , Antígenos Virais/imunologia , Células Clonais , Glutationa Transferase/genética , HIV-1/imunologia , Humanos , Modelos Imunológicos , Peptídeos/imunologia , Receptores de Antígenos de Linfócitos T alfa-beta/imunologia , Proteínas Recombinantes/imunologia , Vacinas/genética , Vacinas/imunologiaRESUMO
Immunodominance or cripticity of a peptide-borne determinant may be influenced by the protein context in which the epitope is embedded. In this frame, we previously showed that certain human T cell clones, derived from different donors, may differentially recognize the RT248-262 helper determinant depending on whether it is provided to the presenting cells as a synthetic peptide or as a recombinant carrier protein to which the sequence of interest is fused. We now report that, upon in vitro immunization of human PBL with autologous APC, the epitope-specific TCRVB repertoire obtained when selection is applied by pulsing the APC with the cognate synthetic peptide is different from that found when a recombinant protein is used in which the antigenic sequence is placed at either a N-terminal or C-terminal location of the GST carrier. As the TCRVB distribution is not a function of the APC used, we propose that processing of different recombinant molecules containing the same epitope may generate MHC/peptide complexes which, being antigenically diverse, may recruit distinct TCR specificities. These findings may be relevant for evaluating and predicting the immunogenic potential of subunit vaccines based on synthetic peptides or on recombinant proteins as compared to the native antigen.
Assuntos
Antígenos HIV/imunologia , Transcriptase Reversa do HIV/imunologia , HIV-1/enzimologia , Receptores de Antígenos de Linfócitos T alfa-beta/imunologia , Sequência de Aminoácidos , Linhagem Celular , HIV-1/imunologia , Humanos , Dados de Sequência Molecular , Proteínas Recombinantes de Fusão/imunologiaRESUMO
The intestinal mucosal lesion in celiac disease is characterized by a predominant T-cell infiltration of both epithelium and lamina propria. However, a restricted use of T-cell receptors (TCR) in T lymphocytes infiltrating the jejunal mucosa of celiac patients has not been reported. Based on an immunohistochemical survey of jejunal biopsies from a cohort of untreated celiac patients, we demonstrated a small but significant increase of V beta 8.1/2+ T cells in the lamina propria, but not in the epithelium nor in the peripheral blood. Sequence analysis indicated the existence of a variable degree of clonality of V beta 8+ T cells in the celiac mucosa. More importantly, the recurrence of identical CDR3 regions in some patients was also observed. The altered distribution of V beta 8+ T cells and the presence of identical CDR3 regions in celiac patients, but not in controls was independently confirmed by CDR3 size analysis in a further cohort of patients. These findings suggest that disease-specific variations of the TCRBV8 repertoire are present in the small intestinal mucosa of untreated celiac patients.
Assuntos
Doença Celíaca/genética , Rearranjo Gênico da Cadeia beta dos Receptores de Antígenos dos Linfócitos T , Mucosa Intestinal/imunologia , Jejuno/imunologia , Receptores de Antígenos de Linfócitos T alfa-beta/química , Linfócitos T/imunologia , Sequência de Aminoácidos , Sequência de Bases , Doença Celíaca/imunologia , Doença Celíaca/patologia , Técnica Indireta de Fluorescência para Anticorpo , Teste de Histocompatibilidade , Humanos , Mucosa Intestinal/patologia , Jejuno/patologia , Dados de Sequência Molecular , Receptores de Antígenos de Linfócitos T alfa-beta/imunologiaRESUMO
N- and C-terminal extensions of naturally processed MHC class II-bound peptides may affect TCR recognition. In fact, residues immediately flanking the minimal epitope on either side can contact the MHC groove and modify the interaction with a TCR. We report now that residues much farther away from the peptide core can also modulate TCR recognition in a functional antigen presentation system. To show this, we isolated from the same donor DR5-restricted T cell clones, specific for the HIV-1 RT(248-262) sequence and differing in their ability to respond to recombinant antigens obtained by insertion of the epitope in different positions of schistosomal, human, or murine glutathione-S-transferase (GST). We found that the reactivity profile of individual clones was related to their TCR fine specificity, suggesting that processing can generate determinants focused onto the same epitope, but antigenically distinct. In addition, we analyzed the response of this panel of T-helper cell clones against GST-derived recombinant antigens in which the epitope was flanked by stretches of polyalanine or polyserine on either side. These spacers had different effects on TCR recognition suggesting that secondary structures outside the core peptide may influence MHC/epitope complex recognition over a distance of 15-30 residues from the determinant.
Assuntos
Epitopos de Linfócito T/imunologia , Transcriptase Reversa do HIV/imunologia , Antígenos de Histocompatibilidade Classe II/imunologia , Peptídeos/imunologia , Processamento de Proteína Pós-Traducional , Receptores de Antígenos de Linfócitos T/imunologia , Sequência de Aminoácidos , Aminoácidos , Animais , Apresentação de Antígeno/imunologia , Separação Celular , Glutationa Transferase/genética , Glutationa Transferase/imunologia , Humanos , Camundongos , Dados de Sequência Molecular , Mutagênese InsercionalRESUMO
We report that a subset of circulating cells reacting with a monoclonal antibody raised against a protein marker is significantly increased in the peripheral blood of women carrying benign or malignant breast diseases, particularly in patients under 55 years of age with ductal mammary carcinomas. These cells were statistically (confidence level of 99%) less represented in a control population including healthy women or women carrying carcinomas of origin other than breast. Double staining analysis showed that they harbor markers of dendritic cells and exhibit endo- cytic activity, as determined by their ability to internalize FITC-dextran particles. Their dendritic morphology was further demonstrated by electron microscopy of sorted antibody-positive cells. However, expression of surface molecules, such as CD34 and CD14, usually not present in differentiated populations of dendritic cells was also observed. Adherent cells of patients with breast ductal carcinoma including mostly cells of this new subset were efficient stimulators of mixed lymphocyte reaction, attaining maximal stimulatory activity attained after TNFalpha treatment. In conclusion, we have shown that a subset of cells characterized by a phenotype suggestive of a yet undescribed stage of maturation of the dendritic cell lineage is accumulated in the blood of patients affected by breast proliferative disorders.
Assuntos
Neoplasias da Mama/imunologia , Carcinoma Ductal de Mama/imunologia , Células Dendríticas/imunologia , Adulto , Anticorpos Monoclonais/imunologia , Neoplasias da Mama/sangue , Neoplasias da Mama/ultraestrutura , Carcinoma Ductal de Mama/sangue , Carcinoma Ductal de Mama/ultraestrutura , Células Dendríticas/classificação , Células Dendríticas/ultraestrutura , Endocitose/imunologia , Feminino , Sangue Fetal/citologia , Sangue Fetal/imunologia , Humanos , Imunofenotipagem , Leucócitos Mononucleares/classificação , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/ultraestrutura , Microscopia Eletrônica , Pessoa de Meia-Idade , Coloração e Rotulagem/métodosRESUMO
The general biological properties of T cell related lymphokines are reviewed here. In particular the stimulatory function of IL-3 on a subset of T cells isolated from the peripheral blood of a normal individual was demonstrated. These cells were CD4 and CD8 negative and expressed the alpha/beta T cell receptor instead of the expected gamma/delta heterodimer. Previously IL-3 has been described to induce the growth of hemopoietic cells, chiefly progenitor cells and mast cells. Moreover IL-7, a newly cloned lymphokine, was found to support the growth and expansion of the human nature T cells. Either T cell clones phenotype and function, or resting T cells were stimulated by IL-7. Finally, the lymphokine production by the T cell clones which have or not cytotoxic capacity was analysed. Different amounts of mRNA transcripts and translated proteins were produced by the two types of T cell clones. This could represent an important tool with which to classify T cells having a function unrelated to the membrane phenotype.
Assuntos
Linfocinas/fisiologia , Subpopulações de Linfócitos T/fisiologia , Animais , Divisão Celular/efeitos dos fármacos , Citotoxicidade Imunológica , Regulação da Expressão Gênica/efeitos dos fármacos , Granzimas , Células-Tronco Hematopoéticas/efeitos dos fármacos , Humanos , Interleucina-3/farmacologia , Linfocinas/farmacologia , Camundongos , RNA Mensageiro/análise , Receptores de Antígenos de Linfócitos T/análise , Serina Endopeptidases/metabolismo , Subpopulações de Linfócitos T/efeitos dos fármacos , Linfócitos T Citotóxicos/metabolismoRESUMO
The immune system is able to respond more vigorously to the second contact with a given antigen than to the first contact. Vaccination protocols generally include at least two doses, in order to obtain high antibody titers. We want to analyze the relation between the time elapsed from the first dose (priming) and the second dose (boost) on the antibody titers. In this paper, we couple in vivo experiments with computer simulations to assess the effect of delaying the second injection. We observe that an interval of several weeks between the prime and the boost is necessary to obtain optimal antibody responses.
Assuntos
Sistema Imunitário/fisiologia , Algoritmos , Doença de Alzheimer/imunologia , Animais , Anticorpos/química , Formação de Anticorpos , Antígenos/química , Simulação por Computador , Ensaio de Imunoadsorção Enzimática/métodos , Humanos , Sistema Imunitário/imunologia , Memória Imunológica , Macrófagos/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Transgênicos , Software , Vacinação , Vacinas de Partículas Semelhantes a VírusAssuntos
Autoimunidade/imunologia , Antígenos de Histocompatibilidade Classe II/imunologia , Modelos Animais , Linfócitos T/imunologia , Animais , Complexo Antígeno-Anticorpo/imunologia , Epitopos/imunologia , Glutationa Transferase/imunologia , Haplótipos , Humanos , Camundongos , Proteínas Recombinantes/imunologiaAssuntos
Citotoxicidade Imunológica , Antígenos HIV/imunologia , HIV-1/imunologia , Vacinas contra a AIDS/imunologia , Animais , Bacteriófagos/genética , Bacteriófagos/imunologia , Epitopos , Antígeno HLA-A2/genética , Humanos , Técnicas In Vitro , Camundongos , Camundongos Transgênicos , Biblioteca de Peptídeos , Linfócitos T Citotóxicos/imunologia , Linfócitos T Auxiliares-Indutores/imunologiaAssuntos
Doenças Autoimunes/etiologia , Antígenos HLA-DP/biossíntese , Antígenos HLA-DQ/biossíntese , Antígenos HLA-DR/biossíntese , Ilhotas Pancreáticas/metabolismo , Glândula Tireoide/metabolismo , Doenças Autoimunes/imunologia , Células Epiteliais , Epitélio/imunologia , Epitélio/metabolismo , Humanos , Ilhotas Pancreáticas/imunologia , Glândula Tireoide/citologia , Glândula Tireoide/imunologiaRESUMO
CD4 and CD8 T-cell clones were generated using the Mx9 monoclonal antibody (mAb), which recognizes the V beta 8 T-cell receptor (TcR) gene family. The interaction of these clones with the Mx9 antibody was analysed and all were found to be specifically stimulated to proliferate by plastic-adherent Mx9. In the presence of Mx9 or its F(ab')2 fragment, CD8+ Mx9+ clones were capable of specifically lysing the CD4+ Mx9+ T-cell clones. No lysis was seen of Mx9- T cells, or in the absence of the antibody. Conversely CD4+ Mx9+ T cells did not have lytic function. These results indicate that cross-linking of T cells via their antigen-specific receptors may initiate a unidirectional killing. Unlike previously reported lytic systems involving anti-TcR antibodies (e.g. anti-CD3), these results suggest that this mechanism may have an important physiological role in immune regulation. Anti-idiotypic antibodies have been shown to recognize T-cell receptors. These may exert profound immunosuppressive effects by inducing the lysis of the helper cells or B cells.
Assuntos
Citotoxicidade Imunológica , Receptores de Antígenos de Linfócitos T/imunologia , Linfócitos T/imunologia , Divisão Celular , Separação Celular , Células Clonais/citologia , Células Clonais/imunologia , Citometria de Fluxo , Humanos , Idiótipos de Imunoglobulinas/imunologia , Linfócitos T/citologiaRESUMO
The analysis of T lymphocytes infiltrating tissues afflicted by autoimmune diseases may provide major clues towards understanding the pathogenesis of such diseases. Currently the best approach to studying heterogeneous populations such as T lymphocytes involves long-term culture and cloning. In order to grow and clone T lymphocytes, regular restimulation with the specific antigen is essential, otherwise growth will stop and/or specificity may be lost. In autoimmune diseases the antigens involved in triggering the immunological reaction of T cells are usually unknown. Therefore an alternative way of stimulating T lymphocytes without loss of specificity is clearly needed. Here we describe the cloning and expansion of antigen-specific T cell clones from the blood of a healthy donor to sizeable numbers of cells (greater than 10(8)) by means of anti-CD3 monoclonal antibody and recombinant IL-2. The results obtained showed that this approach can be used to clone and 'expand' T lymphocytes that retain antigen specificity over a prolonged period, in this case over 10 weeks. This technique has been used to clone and expand T lymphocytes infiltrating the affected tissues in a variety of autoimmune disorders such as Hashimoto's thyroiditis, Graves' disease, and rheumatoid arthritis, and is an efficient method of propagating T cells, by mimicking the antigenic stimulus.
Assuntos
Anticorpos Monoclonais/imunologia , Células Apresentadoras de Antígenos/imunologia , Interleucina-2/farmacologia , Linfócitos T/citologia , Antígenos de Diferenciação de Linfócitos T/análise , Antígenos Virais/imunologia , Artrite Reumatoide/patologia , Doenças Autoimunes/patologia , Células Cultivadas , Células Clonais , Humanos , Vírus da Influenza A/imunologia , Linfócitos T/patologia , Tireoidite Autoimune/patologiaRESUMO
We performed limiting dilution culture of T cells from a patient affected by primary immunodeficiency as a result of complete lack of adenosine deaminase (ADA) activity and also affected by insulin-dependent diabetes mellitus (type I diabetes). Despite the occurrence of immunodeficiency, we were able to raise and grow T cell clones derived from this patient in long-term culture. These T cells displayed ADA enzymatic activity and produced interleukin-2 after engagement of their T cell receptor (TCR)/CD3 complex. We analyzed the TCR repertoire of such clones by nucleotide sequencing of TCR beta chains. The results show that the T cell clones express different V beta but similar J regions. However, the CDR3 regions which are implicated in antigen recognition were found to be heterogeneous.