Decline of T cell-related immune functions in cancer patients and an attempt to restore them through infusion of activated autologous T cells.

We developed a scoring system that can combine several immunological parameters and express the immune status of individuals as a simple numeral. T cell immune score was obtained by using 5T cell-related parameters: number of T cells, ratio of CD4(+)T cells to CD8(+)T cells, number of naïve T cells, ratio of naïve T cells to memory T cells, and T cell proliferative index (TCPI). TCPI was calculated by using number of T cells and their proliferative activity. We assessed T cell immune score in 103 patients with colorectal cancer and 51 healthy age-matched controls. The results were as follows: (1) T cell-immune score of patients in stages I-IV before surgery was significantly decreased as compared with controls. (2) The number of regulatory T cells in patients in stages I-IV gradually increased with disease progression. (3) T cell immune score was strongly suppressed after surgery, but were recovered to the initial level within a month. (4) Furthermore, restoration of immunological function was attempted in cancer patients by infusion of activated autologous T cells. The effectiveness was confirmed by an increase of TCPI in many cancer patients.

Dok-1 and Dok-2 are negative regulators of T cell receptor signaling.

Interaction of the TCR complex with self- or foreign peptides is a central event in the immune responses. Upon TCR stimulation, a protein-tyrosine kinase (PTK), ZAP-70, is recruited to signaling units of the TCR complex, such as TCRzeta, to play an essential role in T cell activation. Here, we find that mice lacking adaptor proteins Dok-1 and Dok-2 show augmented responses to thymus-dependent, but not thymus-independent, antigens, and that their T cells show elevated responses to TCR stimulation, including the activation of ZAP-70 and subsequent proliferation and cytokine production. Furthermore, the forced expression of Dok-1 or Dok-2 in a CD3(+)CD4(+) T cell clone inhibited the activation of ZAP-70 upon TCR stimulation. Interestingly, the Dok-1 and Dok-2 COOH-terminal moieties bearing the src homology 2 target motifs were dispensable for this negative regulation, even though they are crucial for the known adaptor function of Dok-family proteins. Thus, by an as yet unidentified mechanism, Dok-1 and Dok-2 play an essential role in the negative regulation of TCR signaling. Consistently, all mice lacking these proteins exhibited elevated titers of antibodies to double-stranded DNA and developed lupus-like renal disease.

Quantitative analysis of antigen specific IgE in tears in comparison to serum samples.

We determined pollen specific IgE in tears and compared these results to the concentration of specific IgE in serum samples. We obtained tears (using Schirmer strips) and serum samples from subjects with Japanese cedar (Cryptomeria japonica) pollinosis, and tested for C. japonica pollen specific IgE using a quantitative ELISA. Time kinetic analyses through the pollen season showed that specific IgE levels in tears were found to increase earlier than those in sera and reached their maximum at the end of or after the pollen season, from March to early June. In the C. japonica pollen free season, July to December, the specific IgE levels in tears decreased, although the serum levels remained relatively high. These results indicate that the quantitative assay for specific IgE in tears might be useful to identify specific eye allergens.

Tumor necrosis factor-alpha inhibition reduces CXCL-8 levels but fails to prevent fibrin generation and does not improve outcome in a rabbit model of endotoxic shock.

The effects of a monoclonal antibody (mAb) to tumor necrosis factor-alpha (TNF-alpha) were examined in a rabbit model of endotoxic shock. Intravenous administration of lipopolysaccharide (100 microg/kg/hr) for 6 hours (n = 11) increased TNF-alpha levels. Fibrinogen was partially consumed, and fibrin deposits were seen in kidney and lungs at 24 hours. Mortality at 24 hours was 64%. Levels of interleukin-8 (aka CXCL-8) were notably increased. Mean arterial pressure (MAP) and leukocyte counts decreased, whereas creatinine levels were enhanced. The anti-TNF-alpha mAb (20 mg/kg i.v. bolus + 5 mg/kg/h i.v. for the first 90 minutes) (n = 10) efficiently inhibited the TNF-activity. Rabbits exhibited lower CXCL-8 levels; MAP improved, the decrease in leukocyte counts was partially prevented and creatinine levels were lower, but fibrinogen, fibrin deposits in kidneys and lungs and mortality, 55%, were similar to the LPS group. Rabbits that did not survive exhibited lower fibrinogen levels, more fibrin in kidneys and lungs and higher CXCL-8 and creatinine levels than survivors, while there were no differences in TNF-alpha, MAP and leukocytes. Thus, the inhibition of TNF-alpha, although beneficial through lowering CXCL-8 levels, is not enough to improve the outcome, which could be partly due to the inability to prevent the fibrin deposits formation in kidneys and lungs.

A highly sensitive quantitative immunochromatography assay for antigen-specific IgE.

We have developed a highly sensitive quantitative enzyme immunochromatography system for antigen-specific IgE, which is clinically important for the diagnosis of allergic diseases. The system uses alkaline phosphatase-labeled anti-IgE antibody (ALP-anti-IgE) and immobilized target antigen, for example cedar pollen antigen, on a membrane. Antigen-specific IgE present in the serum binds the immobilized antigen after complexing with the ALP-anti-IgE. Subsequently, the enzyme substrate migrates to the complex on the antigen line, which is stained blue. The intensity of the staining was analyzed by a quantitative detector, but can also be assessed directly by the naked eye. This system was able to detect 0.2 U/ml of IgE specific for Japanese cedar pollen. The results correlated well (Spearman's rank correlation coefficient, 0.95) to those in AlaSTAT system, which is a reliable enzyme-linked immunosorbent assay (ELISA) method. This antigen-specific IgE assay system is suitable for point-of-care testing.