Colony-stimulating factor-1 suppresses responses to CpG DNA and expression of toll-like receptor 9 but enhances responses to lipopolysaccharide in murine macrophages

During bacterial infections, the balance between resolution of infection and development of sepsis is dependent upon the macrophage response to bacterial products. We show that priming of murine bone marrow-derived macrophages (BMMs) with CSF-1 differentially regulates the response to two such stimuli, LPS and immunostimulatory (CpG) DNA. CSF-1 pretreatment enhanced IL-6, IL-12, and TNF-alpha production in response to LPS but suppressed the same response to CpG DNA. CSF-1 also regulated cytokine gene expression in response to CpG DNA and LPS; CpG DNA-induced IL-12 p40, IL-12 p35, and TNF-alpha mRNAs were all suppressed by CSF-1 pretreatment. CSF-1 pretreatment enhanced LPS-induced IL-12 p40 mRNA but not TNF-alpha and IL-12 p35 mRNAs, suggesting that part of the priming effect is posttranscriptional. CSF-1 pretreatment also suppressed CpG DNA-induced nuclear translocation of NF-kappaB and phosphorylation of the mitogen-activated protein kinases p38 and extracellular signal-related kinases-1/2 in BMMs, indicating that early events in CpG DNA signaling were regulated by CSF-1. Expression of Toll-like receptor (TLR)9, which is necessary for responses to CpG DNA, was markedly suppressed by CSF-1 in both BMMs and thioglycolate-elicited peritoneal macrophages. CSF-1 also down-regulated expression of TLR1, TLR2, and TLR6, but not the LPS receptor, TLR4, or TLR5. Hence, CSF-1 may regulate host responses to pathogens through modulation of TLR expression. Furthermore, these results suggest that CSF-1 and CSF-1R antagonists may enhance the efficacy of CpG DNA in vivo.

Promiscuous CTL recognition of viral epitopes on multiple human leukocyte antigens: Biological validation of the proposed HLA A24 supertype

Multiple HLA class I alleles can bind peptides with common sequence motifs due to structural similarities in the peptide binding cleft, and these groups of alleles have been classified into supertypes. Nine major HLA supertypes have been proposed, including an A24 supertype that includes A*2301, A*2402, and A*3001. Evidence for this A24 supertype is limited to HLA sequence homology and/or similarity in peptide binding motifs for the alleles. To investigate the immunological relevance of this proposed supertype, we have examined two viral epitopes (from EBV and CMV) initially defined as HLA-A*2301-binding peptides. The data clearly demonstrate that each peptide could be recognized by CTL clones in the context of A*2301 or A*2402; thus validating the inclusion of these three alleles within an A24 supertype. Furthermore, CTL responses to the EBV epitope were detectable in both A*2301(+) and A*2402(+) individuals who had been previously exposed to this virus. These data substantiate the biological relevance of the A24 supertype, and the identification of viral epitopes with the capacity to bind promiscuously across this supertype could aid efforts to develop CTL-based vaccines or immunotherapy. The degeneracy in HLA restriction displayed by some T cells in this study also suggests that the dogma of self-MHC restriction needs some refinement to accommodate foreign peptide recognition in the context of multiple supertype alleles.

Provision of 4-1BB ligand enhances effector and memory CTL responses generated by immunization with dendritic cells expressing a human tumor-associated antigen

Up-regulation of receptor-ligand pairs during interaction of an MHC-presented epitope on dendritic cells (DCs) with cognate TCR may amplify, sustain, and drive diversity in the ensuing T cell immune response. Members of the TNF ligand superfamily and the TNFR superfamily contribute to this costimulatory molecule signaling. In this study, we used replication deficient adenoviruses to introduce a model tumor-associated Ag (the E7 oncoprotein of human papillomavirus 16) and the T cell costimulatory molecule 4-IBBL into murine DCs, and monitored the ability of these recombinant DO to elicit E7-directed T cell responses following immunization. Splenocytes from mice immunized with DCs expressing E7 alone elicited E7-directed effector and memory CTL responses. Coexpression of 4-1BBL in these E7-expressing DO increased effector and memory CTL responses when they were used for immunization. 4-1BBL expression up-regulated CD80 and CD86 second signaling molecules in DO. We also report an additive effect of 4-IBBL and receptor activator of NF-kappaB/receptor activator of NF-kappaB ligand coexpression in E7-transduced DC inummogens on E7-directed effector and memory CTL responses and on MHC class II and CD80/86 expression in DCs. Additionally, expression of 4-1BBL in E7-transduced DCs reduced nonspecific T cell activation characteristic of adenovirus vector-associated immunization. The results have generic implications for improved or tumor Ag-expressing DC vaccines by incorporation of exogenous 4-1BBL. There are also specific implications for an improved DC-based vaccine for human papillomavirus 16-associated cervical carcinoma.

Inhibition of early tumor growth requires Jα18-positive (natural killer T) cells

The role of natural killer T (NKT) cells in the immune response to tumor cells has been largely unexplored. As a model of adoptive tumor immunotherapy, cells from the draining lymph nodes of mice immunized with a tumor-specific or irrelevant antigen were transferred to naive recipients with established tumor. Inhibition of early tumor growth (day 4) required the transfer of both CD8(+) and Jalpha18(+) (NKT) cells from immunized animals without regard to immunogen. In contrast, CD8(+) cells, but not Jalpha18(+) cells, were necessary for the inhibition of late tumor growth (day 8). Thus, the developing tumor changes in sensitivity to NKT-mediated events and the role for NKT cells cannot be replaced by the presence of tumor-specific cells during early tumor growth. This suggests that recruitment/activation of Jalpha18(+) NKT cells is an important consideration during the immune therapy of early stage tumors.

IL-10 Mediates Suppression of the CD8 T Cell IFN-γ Response to a Novel Viral Epitope in a Primed Host

Priming to Ag can inhibit subsequent induction of an immune response to a new epitope incorporated into that Ag, a phenomenon referred to as original antigenic sin. In this study, we show that prior immunity to a virus capsid can inhibit subsequent induction of the IFN-gamma effector T cell response to a novel CD8-restricted antigenic epitope associated with the virus capsid. Inhibition does not involve Ab to the virus capsid, as it is observed in animals lacking B cells. CD8-restricted virus-specific T cell responses are not required, as printing to virus without CTL induction is associated with inhibition. However, IL-10(-/-) mice, in contrast to IL-10(+/+) mice, generate CD8 T cell and Ab responses to novel epitopes incorporated into a virus capsid, even when priming to the capsid has resulted in high titer Ab to the capsid. Furthermore, capsid-primed mice, unable to mount a response to a novel epitope in the capsid protein, are nevertheless able to respond to the same novel epitope delivered independently of the capsid. Thus, inhibition of responsiveness to a novel epitope in a virus-primed animal is a consequence of secretion of IL-10 in response to presented Ag, which inhibits local generation of new CD8 IFN-gamma-secreting effector T cells. Induction of virus- or tumor Ag-specific CD8 effector T cells in the partially Ag-primed host may thus be facilitated by local neutralization of IL-10.

Enhanced effector and memory CTL responses generated by incorporation of receptor activator of NF-kappa B(RANK)/RANK ligand costimulatory molecules into dendritic cell immunogens expressing a human tumor-specific antigen

The outcome of dendritic cell (DC) presentation of Ag to T cells via the TCR/MHC synapse is determined by second signaling through CD80/86 and, importantly, by ligation of costimulatory ligands and receptors located at the DC and T cell surfaces. Downstream signaling triggered by costimulatory molecule ligation results in reciprocal DC and T cell activation and survival, which predisposes to enhanced T cell-mediated immune responses. In this study, we used adenoviral vectors to express a model tumor Ag (the E7 oncoprotein of human papillomavirus 16) with or without coexpression of receptor activator of NF-kappaB (RANK)/RANK ligand (RANKL) or CD40/CD40L costimulatory molecules, and used these transgenic DCs to immunize mice for the generation of E7-directed CD8(+) T cell responses. We show that coexpression of RANK/RANKL, but not CD40/CD40L, in E7-expressing DCs augmented E7-specific IFN-gamma-secreting effector and memory T cells and E7-specific CTLs. These responses were also augmented by coexpression of T cell costimulatory molecules (RANKL and CD40L) or DC costimulatory molecules (RANK and CD40) in the E7-expressing DC immunogens. Augmentation of CTL responses correlated with up-regulation of CD80 and CD86 expression in DCs transduced with costimulatory molecules, suggesting a mechanism for enhanced T cell activation/survival. These results have generic implications for improved tumor Ag-expressing DC vaccines, and specific implications for a DC-based vaccine approach for human papillomavirus 16-associated cervical carcinoma.

Força de preensão – Análise de concordância entre dois dinamómetros: JAMAR vs E‑Link

Introdução – Avaliar a força de preensão mostrou ser de primordial importância pela sua relação com a capacidade funcional dos indivíduos, permitindo determinar níveis de risco para incapacidade futura e, assim, estabelecer estratégias de prevenção. Grande parte dos estudos utiliza o dinamómetro hidráulico JAMAR que fornece o valor da força isométrica obtida durante a execução do movimento de preensão palmar. Contudo, existem outros dinamómetros disponíveis, como é o caso do dinamómetro portátil computorizado E‑Link (Biometrics) que fornece o valor da força máxima (peak force), para além de outras variáveis, como a taxa de fadiga. Não existem, contudo, estudos que nos permitam aceitar e comparar ou não os valores obtidos com os dois equipamentos e porventura utilizá‑los indistintamente. Objetivos – Avaliar a concordância entre as medições da força de preensão (força máxima ou peak force em Kg) obtida a partir de dois equipamentos diferentes (dinamómetros portáteis): um computorizado (E‑Link, Biometrics) e outro hidráulico (JAMAR). Metodologia – Foram avaliados 29 indivíduos (13H; 16M; 22±7 anos; 23,2±3,3 kg/m2) em 2 dias consecutivos, na mesma altura do dia. A posição de teste escolhida foi a recomendada pela Associação Americana de Terapeutas Ocupacionais e foi escolhido o melhor resultado de entre 3 tentativas para a mão dominante. Realizou‑se uma análise correlacional entre os valores obtidos na variável analisada em cada equipamento (coeficiente de Spearman) e uma análise de Bland & Altman para verificar a concordância entre as duas medições. Resultados – O coeficiente de correlação entre as duas medições foi elevado (rS= 0,956; p<0,001) e, pela análise de Bland & Altman, os valores obtidos encontram‑se todos dentro do intervalo da média±2SD. Conclusões – As duas medições mostraram ser concordantes, revelando que os dinamómetros testados podem ser comparáveis ou utilizados indistintamente em diferentes estudos e populações. ABSTRACT: Introduction – Assess grip strength has proved to be of vital importance because of its relationship with functional capacity of individuals, in order to determine levels of risk for future disability and thereby establish prevention strategies. Most studies use the JAMAR Hydraulic dynamometer that provides the value of isometric force obtained during the performance of grip movement. However, there are other dynamometers available, such as portable computerized dynamometer E‑Link (Biometrics), which provides the value of maximum force (peak force) in addition to other variables as the rate of fatigue. There are no studies that allow us to accept or not and compare values obtained with both devices and perhaps use them interchangeably. Purpose – To evaluate the agreement between the measurements of grip strength (peak force or maximum force in kg) obtained from two different devices (portable dynamometers): a computerized (E‑Link, Biometrics) and a hydraulic (JAMAR). Methodology – 29 subjects (13H, 16M, 22 ± 7 years, 23.2 ± 3.3 kg/m2) were assessed on two consecutive days at the same time of day. The test position chosen was recommended by the American Association of Occupational Therapists and was considered the best result from three attempts for the dominant hand. A correlation was studied between values obtained in the variable analyzed in each equipment (Spearman coefficient) and Bland‑Altman analysis to assess the agreement between the two measurements. Results – The correlation coefficient between the two measurements was high (rs = 0,956, p <0,001) and Bland & Altman analysis of the values obtained are all within the range of mean±2SD. Conclusions – The two measurements were shown to be concordant, revealing that the tested dynamometers can be comparable or used interchangeably in different studies and populations.

Tilted boxes on inclined planes

We propose the study of a box placed on an inclined plane, with an initial tilt with respect to the plane. This is a paradigmatic example of the role played by friction as a link between translational and rotational motion. This example has two advantages over the usual example of a sphere (or cylinder) rolling down an inclined plane. First, it provides a good model for a much greater variety of "real-life" situations. Second, it exhibits a much richer structure in parameter space, even when the box starts from rest. (C) 2000 American Association of Physics Teachers.

Auto-determinação e inclusão social das pessoas com deficiência mental - uma intervenção centrada no sujeito

Dotar as pessoas com deficiência mental com competências para se autodeterminarem e terem a oportunidade de concretizar a sua plena inclusão social, é um desafio colocado à sociedade actual. Torna-se importante colocar em prática o que diferentes autores e organizações como a American Association of Mental Retardation defendem, criando condições para que os profissionais, famílias e comunidade possam ser os facilitadores deste processo. Neste sentido foi implementado no Centro de Reabilitação de Ponte de Lima um modelo de intervenção específico baseado na promoção e desenvolvimento da autonomia pessoal, social e de realização da pessoa com deficiência mental e criado um instrumento de observação e registo que reflecte essa forma de intervenção designado por Protocolo de Registo e Avaliação de Competências - PRAC. Neste estudo realizou-se uma análise ao instrumento em causa, pretendendo dar um contributo para a sua posterior validação. Nesse sentido, utilizou-se uma metodologia qualitativa e quantitativa para analisar se o instrumento pode ou não ser considerado representativo da capacidade de autodeterminação; se é estável quando utilizado por mais que um utilizador; se descrimina os indivíduos com maior ou menor autonomia e se os itens quando sujeitos à análise factorial, evidenciam os constructos teóricos previamente traçados. Muito embora o PRAC tenha sido pensado e estruturado para pessoas com deficiência mental, neste estudo foi utilizado por um grupo diversificado de profissionais oriundos de áreas distintas o que veio comprovar que o instrumento pode ser utilizado em diferentes contextos e com público-alvo mais alargado. Os resultados evidenciados são consistentes, permitindo respostas positivas às questões elaboradas, é de referir contudo que necessitam de um maior aprofundamento de forma a estabelecer outro tipo de generalizações.

What do we know about the α/β for prostate cancer?

Since last decade, the debate on the parameter which reflects prostate cancer sensitivity to fractionation in a radiotherapy treatment, the α/β, has become extensive. Unlike most tumors, the low labeling indices (LI) and large potential doubling time that characterize the prostate tumor led some authors to consider that it may behave as a late responding tissue. So far, the existing studies with regard to this subject point to a low value of α/β, around 2.7 Gy, which may be considered as a therapeutic gain in relation to surrounding normal tissues by using fewer and larger fractions. The aim of this paper is to review several estimates that have been made in the last few years regarding the prostate cancer α/β both from clinical and experimental data, as well as the set of factors that have potentially influenced these evaluations.