Assuntos
Linfócitos T CD8-Positivos/virologia , Neoplasias/metabolismo , Células-Tronco/virologia , Adulto , Idoso , Células da Medula Óssea/virologia , Antígenos CD28/biossíntese , Feminino , Humanos , Imunoterapia Adotiva/métodos , Selectina L/biossíntese , Masculino , Pessoa de Meia-Idade , Neoplasias/terapia , Receptores CXCR6 , Receptores de Quimiocinas/biossíntese , Receptores Virais/biossíntese , Células-Tronco/metabolismo , Membro 7 da Superfamília de Receptores de Fatores de Necrose Tumoral/biossínteseRESUMO
AIMS: An important consideration in the design of a tumour vaccine is the ability of tumour-specific cytotoxic T lymphocytes (CTL) to recognise unmanipulated tumour cells in vivo. To determine whether B-CLL might use an escape strategy, the current studies compared B-CLL and normal B cell MHC class I expression. METHODS: Flow cytometry, TAP allele PCR and MHC class I PCR were used. RESULTS: While baseline expression of MHC class I did not differ, upregulation of MHC class I expression by B-CLL cells in response to IFN-gamma was reduced. No deletions or mutations of TAP 1 or 2 genes were detected. B-CLL cells upregulated TAP protein expression in response to IFN-gamma. Responsiveness of B-CLL MHC class I mRNA to IFN-gamma was not impaired. CONCLUSIONS: The data suggest that MHC class I molecules might be less stable at the cell surface in B-CLL than normal B cells, as a result of the described release of beta(2)m and beta(2)m-free class I heavy chains from the membrane. This relative MHC class I expression defect of B-CLL cells may reduce their susceptibility to CTL lysis in response to immunotherapeutic approaches.
Assuntos
Antígenos de Histocompatibilidade Classe I/biossíntese , Antígenos de Histocompatibilidade Classe I/imunologia , Interferon gama/farmacologia , Leucemia Linfocítica Crônica de Células B/metabolismo , Membro 2 da Subfamília B de Transportadores de Cassetes de Ligação de ATP , Membro 3 da Subfamília B de Transportadores de Cassetes de Ligação de ATP , Transportadores de Cassetes de Ligação de ATP/biossíntese , Transportadores de Cassetes de Ligação de ATP/imunologia , Idoso , Apresentação de Antígeno/imunologia , Citometria de Fluxo , Antígenos de Histocompatibilidade Classe I/efeitos dos fármacos , Humanos , Interferon gama/imunologia , Leucemia Linfocítica Crônica de Células B/imunologia , Reação em Cadeia da PolimeraseRESUMO
Core binding factor acute myeloid leukemia (CBF AML), with t(8;21)(q22;q22), inv(16)(p13q22) or t(16;16)(p13;q22) and the associated fusion gene transcripts AML1/ETO or CBFbeta/MYH11, has a favourable clinical prognosis although significant numbers of patients still suffer relapse. We examined the prognostic utility of serial bone marrow minimal residual disease (MRD) monitoring by RQ-PCR in a cohort of patients with CBF AML with long term clinical follow-up. Twenty-nine patients were evaluated with a median follow of 34 months. Twelve relapses occurred at a median of 11 months (range 4 - 17) from diagnosis. RQ-PCR levels at diagnosis, post-induction chemotherapy and post-consolidation were not predictive of outcome. However, a >or=1 log(10) rise at any stage in transcript level relative to the level from a remission bone marrow sample correlated with inferior leukemia free survival (LFS) and imminent morphologic relapse (hazard ratio 8.6). Relapses occurred a median of 60 days (range 45 - 272) after a log(10) rise. A >or=1 log(10) rise in transcript levels strongly predicts subsequent morphologic relapse in CBF AML and therefore defines molecular relapse. Our data support a simple RQ-PCR model for prediction of impending relapse which has the potential for widespread clinical applicability. Prospective identification of high risk patients will enable clinical trials to assess the efficacy of treatment initiated at molecular relapse.
Assuntos
Fatores de Ligação ao Core , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/patologia , Neoplasia Residual/diagnóstico , Valor Preditivo dos Testes , RNA Mensageiro/análise , Adolescente , Adulto , Idoso , Exame de Medula Óssea , Criança , Estudos de Coortes , Feminino , Seguimentos , Humanos , Leucemia Mieloide Aguda/diagnóstico , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Prognóstico , RecidivaRESUMO
Immunosuppression following solid organ transplantation results in impaired T-cell immunity and risk of Epstein-Barr virus (EBV)-positive post-transplant lymphoproliferative disorders (PTLD). The B-cell targeting antibody rituximab has efficacy in PTLD. As B cells are the principle reservoir for EBV, we investigated the effect of rituximab on the persistence of EBV-specific CD8(+) T-cell immunity. To avoid the confounding factor of concurrent immunosuppression to prevent transplant rejection, immunity was analysed in non-transplanted lymphoma patients (i.e. a non-PTLD setting). Cytomegalovirus-specific T-cell immunity was assessed as an internal control. Our data demonstrated that circulating B cells were not critical for maintaining EBV-specific T-cell immunity.
Assuntos
Anticorpos Monoclonais/farmacologia , Antineoplásicos/farmacologia , Linfócitos T CD8-Positivos/efeitos dos fármacos , Infecções por Vírus Epstein-Barr/imunologia , Linfoma não Hodgkin/tratamento farmacológico , Idoso , Anticorpos Monoclonais Murinos , Linfócitos T CD8-Positivos/imunologia , Células Cultivadas , Feminino , Humanos , Imunidade Celular/efeitos dos fármacos , Linfoma não Hodgkin/imunologia , Masculino , Pessoa de Meia-Idade , Rituximab , Linfócitos T Citotóxicos/efeitos dos fármacos , Linfócitos T Citotóxicos/imunologiaRESUMO
The Hyper-CVAD chemotherapy regimen is being increasingly applied to a number of haematological malignancies. We assessed the impact of Hyper-CVAD on peripheral blood stem cell (PBSC) yields and examined the optimal timing of PBSC collection when using this regimen. Seventy-four consecutive patients were identified in whom an attempt was made to collect PBSC, usually on recovery from cycle A or B. Where PBSC collection was attempted after cycle 3B, only 18% (3/17) of patients successfully mobilized. Fifty-seven patients were mobilized on recovery from cycle 1B (n = 13), 2A (n = 22), 2B (n = 14) or 3A (n = 8). Compared with cycle 2A, 1B was not superior in achieving the minimum of > or =2 x 10(6)/kg CD34+ cells (100% vs. 77%, p = 0.13), but was superior in terms of total CD34+ yield (21.4 vs. 3.2 x 10(6)/kg, p < 0.001), achieving the target CD34+ cell count of > or =5 x 10(6)/kg (92% vs 36%, p = 0.002), and obtaining both a minimum (92% vs. 18%, p < 0.001) and target (77% vs. 0%, p < 0.001) graft with a single apheresis. There were no significant differences in PBSC yields following cycles 2A, 2B and 3A. Hyper-CVAD has substantial stem cell toxicity which can be readily circumvented by using the early chemotherapy cycles for mobilization.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Sobrevivência de Enxerto/efeitos dos fármacos , Neoplasias Hematológicas/terapia , Mobilização de Células-Tronco Hematopoéticas , Transplante de Células-Tronco de Sangue Periférico , Adolescente , Adulto , Idoso , Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Remoção de Componentes Sanguíneos/métodos , Ciclofosfamida/administração & dosagem , Ciclofosfamida/efeitos adversos , Dexametasona/administração & dosagem , Dexametasona/efeitos adversos , Intervalo Livre de Doença , Doxorrubicina/administração & dosagem , Doxorrubicina/efeitos adversos , Feminino , Neoplasias Hematológicas/mortalidade , Humanos , Masculino , Pessoa de Meia-Idade , Transplante Autólogo , Vincristina/administração & dosagem , Vincristina/efeitos adversosRESUMO
There are a number of observations that suggest the dsRNA-activated protein kinase, PKR, may play an active role in formation and maintenance of leukemia, including nonrandom chromosomal deletions in acute leukemia as well as truncations and deletions of the PKR gene in some leukemia cell lines. However, there is little direct evidence from patient material that this is so. Here we show that full-length PKR is present but not active in 21 of 28 patient samples from B-cell chronic lymphocytic leukemia (B-CLL). PKR from these patients was unable to auto-activate or phosphorylate substrates but was able to bind dsRNA. Furthermore, the lack of PKR activation was not due to differing levels of the PKR activator, PACT nor of the PKR inhibitor, p58(IPK). We compared PKR status with clinical parameters and disease staging. No differences were found between the 2 groups in terms of staging (modified Rai or Binet), age, CD38 status, p53 status, 11q23 deletion status or CEP12 deletion status. However, there was a significant correlation between deletion in 13q14.3 and lack of PKR activity. We show that B-CLL cells appear to contain a soluble inhibitor of PKR, as lysates from cells lacking PKR activity were able to inhibit exogenous PKR in mixing experiments. Finally, we show suppression of PKR activity was still present following ultrafilitration through a 10,000 Da cutoff filter but was lost upon extraction with phenol/chloroform or by high salt washing. This data suggests loss of PKR activity may contribute to the formation and/or maintenance of CLL.
Assuntos
Leucemia Linfocítica Crônica de Células B/enzimologia , RNA de Cadeia Dupla/metabolismo , eIF-2 Quinase/metabolismo , ADP-Ribosil Ciclase/metabolismo , ADP-Ribosil Ciclase 1 , Idoso , Antígenos CD/metabolismo , Cromossomos Humanos Par 11/genética , Cromossomos Humanos Par 13/genética , Regulação para Baixo , Ativação Enzimática/efeitos dos fármacos , Feminino , Deleção de Genes , Proteínas de Choque Térmico HSP40 , Humanos , Leucemia Linfocítica Crônica de Células B/patologia , Masculino , Glicoproteínas de Membrana , Fosforilação/efeitos dos fármacos , Poli I-C , Proteínas de Ligação a RNA/farmacologia , Proteínas Repressoras/farmacologia , Células Tumorais Cultivadas , Proteína Supressora de Tumor p53/metabolismo , eIF-2 Quinase/antagonistas & inibidores , eIF-2 Quinase/genéticaRESUMO
Re-treatment with rituximab for B-cell non-Hodgkin's lymphoma (NHL) relapsing after previous rituximab therapy has recently been shown to be clinically efficacious. Although the mechanism of resistance to rituximab re-treatment in non-responding patients is unknown, it is possible that loss of CD20 expression in the relapsed NHL could be important in some patients. We examined the incidence and nature of CD20 negative relapses following rituximab therapy in aggressive B-cell NHL treated at our institution. Of a total of 18 patients who received rituximab, 13 have relapsed, with 10 patients subsequently undergoing repeat tissue biopsy. Six of these 10 patients (60%) were shown to have lost CD20 expression by either immunohistochemistry and/or flow cytometry. Furthermore, three of the six patients who relapsed with CD20-negative NHL also suffered relapses at unusual anatomical sites. We conclude that loss of CD20 expression in aggressive B-cell NHL relapsing post-rituximab therapy is common. As such, repeat tissue biopsy should be undertaken to document CD20 expression by both flow cytometry and immunohistochemistry prior to considering repeated courses of rituximab in relapsed aggressive lymphomas.
Assuntos
Anticorpos Monoclonais/uso terapêutico , Antígenos CD20/análise , Antineoplásicos/uso terapêutico , Linfoma de Células B/imunologia , Recidiva Local de Neoplasia/imunologia , Adulto , Idoso , Anticorpos Monoclonais Murinos , Antígenos CD20/imunologia , Feminino , Citometria de Fluxo , Humanos , Imuno-Histoquímica , Incidência , Linfoma de Células B/tratamento farmacológico , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/tratamento farmacológico , Estudos Retrospectivos , RituximabRESUMO
Metastatic melanoma is poorly responsive to treatment, and immunotherapeutic approaches are potentially beneficial. Predictors of clinical response are needed to identify suitable patients. We sought factors associated with melanoma-specific clinical response following intradermal vaccination with autologous melanoma peptide and particulate hepatitis B antigen (HBsAg)-exposed immature monocyte-derived dendritic cells (MDDC). Nineteen patients with metastatic melanoma received a maximum of 8, 2-weekly vaccinations of DC, exposed to HBsAg in addition to autologous melanoma peptides. A further 3 patients received an otherwise identical vaccine that did not include HBsAg. Patients were assessed 1-2 monthly for safety, disease volume, and cellular responses to HBsAg and melanoma peptide. There was no significant toxicity. Of 19 patients receiving HBsAg-exposed DC, 9 primed or boosted a cellular response to HBsAg, and 10 showed no HBsAg response. HBsAg-specific responses were associated with in vitro T cell responses to melanoma peptides and to phytohemagglutinin (PHA). Zero out of 10 non-HBsAg-responding and 4/9 HBsAg-responding patients achieved objective melanoma-specific clinical responses or disease stabilization - 1 complete and 2 partial responses and 1 case of stable disease ( P=0.018). Development of melanoma-specific cellular immunity and T cell responsiveness to mitogen were greater in the group of patients responding to HBsAg. Therefore stimulation of an immune response to nominal particulate antigen was necessary when presented by melanoma peptide-exposed immature DC, to achieve clinical responses in metastatic melanoma. Since general immune competence may be a determinant of treatment response, it should be assessed in future trials on DC immunotherapy.