anti-T Antibodies Research Articles

Sparrows (Passer domesticus L.) as intermediary hosts of Toxoplasma gondii in poultry farms from the "agreste" region of Pernambuco, Brazil

This paper aimed to identify Toxoplasma gondii infection in house sparrows (Passer domesticus, Linneaus 1758) coming from poultry farms in the "agreste" region of the Brazilian state of Pernambuco. 151 sparrows (Passer domesticus) captured in eight broiler, egg layer and commercial laying poultry farms, were used. Indirect hemagglutination test was used to research anti-T. gondii antibodies. Animals that presented titration of 1:16 were destined to DNA research through Polymerase Chain Reaction (PCR) technique, followed by Nested-PCR. It was observed that, from 151 analyzed samples. 91 (60.3%) were reagents and 60 (39.7%) were not reagents. It was verified, through analysis of the distribution of infected animals frequency per farm, that in only one farm (12.5%) no animal reagent to T. gondii was captured. It was also observed that three (30.00%) of the ten samples destined to DNA research for T. gondii were positive to PCR and four (40.00%) were positive to Nested-PCR. Anti-T gondii antibodies occurrence and the molecular identification of the agent confirmed natural T. gondii infection in sparrows from poultry farms in Brazil. Other studies must be carried out to highlight the real importance of these animals in the epidemiological chain and their efficiency in the transmission of the parasite to felines. Therefore, researches that use parasite isolation and molecular techniques to determine genomic profile of the agent present in these poultry farms are needed.

Antigen-dependent T lymphocytes (TcRβ +) are primarily differentiated in the thymus rather than in other lymphoid tissues in sea bass ( Dicentrarchus labrax, L.)

All jawed vertebrates share lymphocyte receptors that allow the recognition of pathogens and the discrimination between self and non-self antigens. The T cell transmembrane receptor (TcR) has a central role in the maturation and function of T lymphocytes in vertebrates via an important role in positive selection of the variable region of TcR αβ/γδ chains. In this study, the TcRβ transcript expression and TcRβ + cell distribution during the ontogeny of the immune system of sea bass ( Dicentrarchus labrax, L.) were analysed. RT-PCR analysis of larvae during early development demonstrated that the β chain transcript is expressed by 19 days post-fertilisation (p.f.). RNA probes specific for the β chain were synthesised and used for in situ hybridisation experiments on 30 day p.f. to 180 day old juvenile larvae. A parallel immunohistochemical study was performed using the anti-T cell monoclonal antibody DLT15 developed in our laboratory [Scapigliati et al., Fish Shellfish Immunol 1996; 6:383–401]. The first thymus anlage was detectable at 32–33 days p.f. (Corresponding to about 27 days post-hatch). DLT15 + cells were detected at day 35 p.f. in the thymus whereas TcRβ + cells were recognisable at day 38 p.f. in the thymus and at day 41 p.f. in the gut. TcRβ + cells were observed in capillaries from 41 to 80 days p.f. At day 46 p.f., TcRβ + cells were identified in the head kidney and were detected in the spleen 4 days later. The present results demonstrate that TcRβ + cells can be differentiated first in the thymus and then in other organs/tissues, suggesting potential TcRβ + cell colonisation from the thymus to the middle gut. Once the epithelial architecture of the thymus is completed with the formation of the cortical-medullary border (around 70–75 days p.f.), DLT15 + cells or TcRβ + cells are confined mainly to the cortex and cortical-medullary border. In particular, a large influx of TcRβ + cells was observed at the cortical-medullary border from 72 to 90 days p.f., suggesting a role in positive selection for this thymic region during the ontogeny of the fish immune system. This study provides novel information about the primary differentiation and distribution of TcRβ + cells in sea bass larvae and juveniles.

Direct Construction of an Open-Sandwich Enzyme Immunoassay for One-Step Noncompetitive Detection of Thyroid Hormone T4

To establish a sensitive noncompetitive immunoassay for thyroxine (T4), we attempted to isolate anti-T4 antibodies from a phage display library based on a phagemid pDong1 ( Dong et al. Anal. Biochem.2009, 36, 386 ), which was designed to enable open-sandwich enzyme-linked immunosorbent assay (OS-ELISA) after selection on immobilized antigen. After the Fab-displaying phage library made from the splenocytes of T4-KLH immunized mice was subjected to biopanning on T4-BSA, two T4-specific clones were obtained. When they were assayed by indirect competitive ELISA, both clones showed low IC(50) (5-13 ng/mL), indicating their high affinity to T4. When they were used for OS-ELISA that detects antigen-dependency of the interaction between variable domains V(H) and V(L), a clone successfully detected 1 ng/mL of T4 with a working range superior to that of competitive IA. OS-ELISA was also performed with maltose binding protein (MBP)-fused V(H)/V(L) of this clone, which showed a detection limit less than 0.1 ng/mL T4. Moreover, the assay showed cross-reactivity with T3 similar to that of competitive ELISA, and also gave a reasonable total serum T4 concentration (90 ng/mL) from ethanol-extracted sample serum using the recombinant proteins. This is the first direct construction of an OS-ELISA system bypassing hybridoma, which will be applicable to the detection of many other small molecule antigens.

Diphtheria and Tetanus Antibody Persistence in Indian Pre-school Children and Response to a Booster Dose of DT Vaccine.

Despite effective vaccines, diphtheria (D) resurged recently in the former socialistic block, and tetanus (T) still occurs in less privileged countries. We studied the antibody persistence for D and T in Indian pre-school children who had received four doses of DTP vaccine and subsequently, the response to a booster dose. Anti-D and anti-T IgG antibodies prior to and one month after a DT vaccine were measured by ELISA in 223 healthy children of 4-6 years who had previously received four doses of the triple vaccine. Adverse reactions were monitored for one month. While 30% and 14% of subjects were susceptible to D and T, respectively, 98% and 100% of them attained seroprotection post-vaccination. Both responses were significant. Local, but not systemic reactions except fever were rather common. A high proportion of the Indian pre-school population is susceptible to D and T, despite of receiving four doses. The current policy of giving the fifth dose at this age is appropriate.

DEVELOPMENT AND VALIDATION OF A TWO-STEP FREE THYROXINE RADIOIMMUNOASSAY BASED ON ANTIBODY COATED TUBES

We describe a two-step radioimmunoassay procedure (RIA) for the measurement of free thyroxine (fT4) in human serum. A commercial antibody with the affinity of 5.5 × 109 L/M was used for this study. The anti-T4 antibody was immobilized on the inner walls of the polystyrene tubes by passive adsorption through normal rabbit IgG and anti-rabbit IgG as immunobridges. The method developed uses very small amounts of antibody with minimum dilution of the sample. The assay covers the range of 0 to 87.5 pmol/L with sensitivity of 0.9 pmol/L. The intra-assay precisions were 6.3% and 7% at 6.7 and 20.7 pmol/L, respectively, for 15 replicates. The inter-assay precision at 6.4 and 24.4 pmol/L were 13% and 11.6%, respectively. The normal range established by analyzing 54 healthy volunteers was 15.3–24.7 pmol/L and that of 69 ambulatory subjects was 14.8–23.3 pmol/L. Progressive dilution of euthyroid and hypothyroid samples up to 50-fold yielded virtually constant values, whereas a registered decrease in free T4 was observed with hyperthyroid samples. Free T4 estimated for 75 samples, including samples from 16 pregnant female subjects, correlated well with the values obtained by a reputed commercial kit with the linear correlation coefficient of 0.84.

Micro Open-Sandwich ELISA to Rapidly Evaluate Thyroid Hormone Concentration from Serum Samples

Thyroxine (T4) is the most commonly measured thyroid hormone for the diagnosis of thyroid function. To elucidate a rapid and sensitive assay for T4, we made a microfluidics-based noncompetitive immunodetection chip system using anti-T4 antibody fragments obtained from a phage display library. Based on the open-sandwich ELISA principle that detects antigen-dependency of the interaction between the two antibody variable regions V(H) and V(L), we could detect less than 1 ng/ml of T4. The assay was also successfully applied to evaluate total T4 concentration in the serum of healthy individuals. This would be the first micro open-sandwich ELISA constructed with antibody fragments directly selected from immunized mice. The system will be applied to the sensitive detection of many diagnostic markers.

Role of dihydropyrimidine dehydrogenase and thymidylate synthase expression in immunohistochemistry of intrahepatic cholangiocarcinoma

Dihydropyrimidine dehydrogenase (DPD) and thymidylate synthase (TS) are key enzymes in the metabolism of 5-fluorouracil and have been implicated as possible prognostic markers for cancer patients. However, the clinical roles of DPD and TS in intrahepatic cholangiocarcinoma (IHCC) have not been investigated. The aim of this study was to clarify the clinicopathological role of DPD and TS expressions in IHCC. Twenty-nine patients who had undergone hepatic resection for IHCC were enrolled in this study. Expressions of DPD and TS in the resected IHCC specimens were examined using anti-DPD or anti-TS antibody. The patients were divided into positive and negative groups according to DPD/TS expressions: DPD-positive group (n = 18) and DPD-negative group (n = 11)/TS-positive group (n = 14) and TS-negative group (n = 15). Clinicopathological factors were compared between the two groups. The overall survival rate was significantly lower in the DPD-negative group than in the DPD-positive group (1-year 36.4% vs. 77.4%, 3-year 18.2% vs. 43.0%; P < 0.05). The disease-free survival rate in the DPD-negative group tended to be lower than that in the DPD-positive group. The overall survival rate or disease-free survival rate did not appear to be associated with the TS-expression status. The Ki-67 labeling index in the DPD-negative group was significantly higher than that in the DPD-positive group (16.9 ± 3.2% vs.13.2 ± 3.3%; P < 0.05). The negative DPD expression was significantly associated with the enhanced tumor cell proliferation and poorer prognosis in patients with IHCC. DPD expression is a potential prognostic indicator for IHCC.

Mycobacterium tuberculosis infection in liver transplantation

Mycobacterium tuberculosis can cause significant infections in liver transplant candidates and recipients. Its nonspecific clinical features and prolonged growth time in culture make the diagnosis difficult, and treating tuberculosis (TB) remains challenging because of significant toxicities and drug-drug interactions. The diagnosis of a latent TB infection may be accomplished with tuberculin skin testing and with the newer interferon-γ release assays, although this infection may be underrecognized because of host factors. Latent TB should be treated, but the degree of liver failure and the likelihood of progression to active TB will dictate whether this should occur before or after transplantation. Patients who have a history of TB, have used muromonab-CD3 or anti-T lymphocyte antibodies, or have experienced allograft rejection or coinfection with cytomegalovirus, Pneumocystis jiroveci, or Nocardia are at the greatest risk of developing active TB. Active TB in transplant patients is difficult to treat because of drug-induced hepatotoxicity and the significant interaction between rifampin and calcineurin inhibitors. In this article, we review the epidemiology, clinical features, and evaluation of transplant candidates and recipients. In addition, we offer recommendations on the appropriate diagnostic and treatment regimens for patients with latent and active TB infections.

Anti‐idiotypic antibody mimicking a T‐antigen‐specific lectin inhibits human epithelial tumor cell proliferation

Cancer-associated mucins show frequent alterations of oligosaccharide chain profile. Terminal structures may be deleted, thereby exposing normally 'cryptic' structures such as Tn (GalNAcα-O-Ser/Thr) and T antigen (Galβ1-3GalNAcα-O-Ser/Thr). Overexpression of these commonly hidden glycoforms, and reduced level of naturally occurring anti-T or anti-Tn antibodies, is associated with epithelial tumor progression and aggressiveness. The lectin from the common edible mushroom Agaricus bisporus (ABL) shows high affinity binding to T antigen, and reversible noncytotoxic inhibitory effect on epithelial tumor cell proliferation. The aim of this study was to induce immune response with tumor-associated glycan specificity and biological activity similar to those of ABL. An anti-idiotypic (Id) antibody strategy was developed using ABL as first template. ABL was purified by affinity chromatography and assayed as immunogen in rabbit. Rabbit IgG was purified from anti-ABL serum using a protein G column, and specific anti-ABL IgG was obtained by affinity chromatography using immobilized ABL. Affinity-purified anti-ABL IgG contained an antibody fraction that recognizes the carbohydrate-binding site of ABL. This IgG was used as immunogen in mouse to yield anti-Id antibody recognizing tumor-associated glycans such as Tn and T antigen. Competitive assays showed that α-anomeric GalNAc is the main binding subsite of anti-Id antibody in glycan recognition. Anti-Id antibody bound human epithelial tumor cells, as shown by cell enzyme-linked immunosorbent assay and immunofluorescence. Anti-Id antibody raised by immunization with affinity-purified anti-ABL IgG had antiproliferative effect on human epithelial tumor cells through apoptosis induction similar to that of ABL. The anti-Id immune response developed here has potential application in cancer therapy.

Cross-sectional survey of Toxoplasma gondii infection in colony cats from urban Florence (Italy).

Cats are the key species in the epidemiology of Toxoplasma gondii infection, even if the proportion of subjects excreting oocysts is low. The aim of the present paper was to obtain information about seroprevalence, oocyst shedding rate and presence of T gondii DNA in faeces collected from an urban population of colony cats in Florence (Tuscany). Fifty European shorthair feral cats were examined for anti-T gondii specific antibodies by a modified agglutination test (MAT), and for oocysts by microscopic examination and for faecal protozoal DNA, by means of a nested polymerase chain reaction (n-PCR) protocol. Twenty-two out of 50 serum samples (44%) were MAT positive. T gondii oocysts were not detected in any of the examined faecal samples. Eight out of 50 faecal specimens (16%) were n-PCR positive and sequencing of the bands was specific for T gondii. Detection by combination of the two methods was higher than single techniques and enhanced the detection of T gondii up to 48%. Our results suggest that the use of MAT plus PCR in faeces may be the best choice for diagnosis of feline toxoplasmosis. Further studies to ascertain the real infectivity of the copro-PCR positive subjects are required.

Pneumococcal Induced T-activation with Resultant Thrombotic Microangiopathy

Thrombotic microangiopathies are disorders resulting from platelet thromboses forming in the microvasculature with resultant schistocyte forms. Hemolytic uremic syndrome (HUS) is a microangiopathic hemolytic anemia often complicated by acute renal failure in children. HUS is typically caused by bacterial infection, most commonly enterohemorrhagic Escherichia coli. Neuraminidase-producing organisms, such as Streptococcus pneumoniae have also been reported as potential etiologies. The pathogenesis in these cases involves cleavage of sialic acid residues from the surfaces of erythrocytes, platelets, and glomerular capillary endothelial cells, exposing the Thomsen-Friedenreich antigen, a process known as T-activation. We describe a 2-year-old girl who presented with pneumococcal pneumonia and sepsis ultimately resulting in a thrombotic microangiopathy with acute renal failure, most consistent with HUS. The patient's direct antiglobulin test was positive. Polyagglutination was observed with human adult serum, but not with umbilical cord serum. Her red blood cells (RBCs) were reactive against peanut and soybean lectins, but not Salvia sclarea or Salvia horminum lectins. These findings are consistent with T-activation. Clinicians should be cognizant of the possibility of T-activation with resultant HUS in patients infected with neuraminidase-producing bacteria. Such patients may be difficult to identify using monoclonal typing antisera, as these typically do not have anti-T antibodies. Whether such patients are at risk for transfusion-associated hemolysis is debatable.

Chemiluminescent immunoassay of thyroxine enhanced by microchip electrophoresis

A homogeneous chemiluminescent immunoassay of thyroxine (T4) enhanced by microchip electrophoresis separation has been developed. The method deployed the competitive immunoreaction of T4 and horseradish peroxidase (HRP)-labeled T4 (HRP–T4) with anti-T4 mouse monoclonal antibody (Ab). HRP–T4 and the HRP–T4–Ab complex were separated and quantified by using microchip electrophoresis (MCE) with chemiluminescence (CL) detection. Highly sensitive CL detection was achieved by means of HPR-catalyzed luminol–H 2O 2 reaction. Due to the effective MCE separation, the CL analytical signal was less prone to sample matrix interference. Under the selected assay conditions, the MCE separation was accomplished within 60 s. The linear range for T4 was 5–250 nM with a detection limit of 2.2 nM (signal/noise ratio = 3). The current method was successfully applied for the quantification of T4 in human serum samples. It was demonstrated that the current MCE–CL-enhanced competitive immunoassay was quick, sensitive, and highly selective. It may serve as a tool for clinical analysis of T4 to assist in the diagnosis of thyroid gland functions.

T4 phages against Escherichia coli diarrhea: Potential and problems

A combination of in vitro and in vivo experiments with comparative phage genomics was used for the rational design of a phage cocktail against E. coli diarrhea. Orally applied T4 coliphages representing three different subgroups (T4-, RB49- and JS98-like phages) had no negative impact on the murine gut microbiota. T4 phages were found with high titers in the cecum and colon and lower titers in the small intestine, but were not detected in the blood, liver or spleen. No adverse effects were observed after one-month exposure to phage nor were serum anti-T4 antibodies detected. T4 phages belonging to the same subgroup showed closely related genomes that differed by 12 (phage JS10 vs. JS98 reference) to 17 (phage JSE vs. RB49 reference) insertion/deletions mostly representing single small ORFs. Bioinformatic analysis did not reveal undesired genes in the T4 genomes. Sequence variability was seen over the tail fibre genes, but the variability did not correlate with phage host range. The investigated T4 phages were not only species- but also strain-specific, necessitating the use of phage cocktails consisting of 10 and 16 T4 phage isolates to cover half to two thirds of E. coli strains representing the five main pathotypes isolated from diarrhea patients.

Tumor-associated Carbohydrate Antigens: Sialyl Lea and T/Tn Antigens in Canine Mammary Tumors

Twenty-eight canine mammary tubulopapillary carcinomas and 14 simple adenomas were studied by immunohistochemistry for the expressions of the tumor-associated carbohydrate antigens. Sialyl Le(a) was detected in 71.42% of the malignant and 92.84% of the benign tumors. Staining with anti-T and anti-Tn monoclonal antibodies revealed that 85.70% of the tubulopapillary carcinomas expressed T and Tn antigens. In contrast, 50% of the adenomas did not express T antigen, and 42.85% of them were only weakly stained for this carbohydrate structure. In the case of Tn antigen, the majority (57.14%) of samples was weakly stained, and no binding was observed in 35.71% of the analyzed specimens. Comparison of average values of reaction intensity (IRS) scale for malignant versus benign tumors by the Mann-Whitney U-test revealed a significant relationship between T and Tn antigens expression and type (malignant vs. benign) mammary tumors. Based on the results obtained, it is suggested that each of the studied antigens can be treated as a tumor-associated antigen of canine mammary tumors. However, only the T and Tn antigens seem to be associated with malignant transformation of mammary gland cells and to be of potential value as diagnostic markers.

Tumor-associated Carbohydrate Antigens: Sialyl Lea and T/Tn Antigens in Canine Mammary Tumors

Twenty-eight canine mammary tubulopapillary carcinomas and 14 simple adenomas were studied by immunohistochemistry for the expressions of the tumor-associated carbohydrate antigens. Sialyl Lea was detected in 71.42% of the malignant and 92.84% of the benign tumors. Staining with anti-T and anti-Tn monoclonal antibodies revealed that 85.70% of the tubulopapillary carcinomas expressed T and Tn antigens. In contrast, 50% of the adenomas did not express T antigen, and 42.85% of them were only weakly stained for this carbohydrate structure. In the case of Tn antigen, the majority (57.14%) of samples was weakly stained, and no binding was observed in 35.71% of the analyzed specimens. Comparison of average values of reaction intensity (IRS) scale for malignant versus benign tumors by the Mann-Whitney U-test revealed a significant relationship between T and Tn antigens expression and type (malignant vs. benign) mammary tumors. Based on the results obtained, it is suggested that each of the studied antigens can be treated as a tumor-associated antigen of canine mammary tumors. However, only the T and Tn antigens seem to be associated with malignant transformation of mammary gland cells and to be of potential value as diagnostic markers.

Preparation of anti-T idiotype monoclonal antibody reacting with human T leukaemic cell lines and with a small percentage of peripheral T lymphocytes

Monoclonal antibody (MoAb) KT38 raised against a human T leukaemic cell line TALL-1, reacted with another T leukaemic cell line Jurkat, but not with any other cell lines tested. The co-modulation of CD3 and KT38 antigen was observed with stimulations of either MoAb T3 or KT38 on both TALL-1 and Jurkat. Upon radioimmunoprecipitation and SDS-PAGE analysis, MoAb KT38 precipitated the heterodimer of 40-60 kD from Jurkat and TALL-1 under reducing conditions. Thus, MoAb KT38 is considered to be an anti-T idiotype (Ti) antibody to TALL-1 and Jurkat cells. MoAb KT38 was also shown to react with a minor population of peripheral blood lymphocytes (PBL) and with very few cells (0.5-2.0%) in the paracortical area of the lymph node. When PBL were stimulated in a KT38-coated culture flask for 5 days, the percentage of KT38-positive PBL was markedly increased. The CD3 antigen on these cultured PBL in the flask was modulated by the stimulation with MoAb KT38. Thus, it is suggested that a common idiotope exists on the T cell receptor of Jurkat, TALL-1 and a small percentage (1.9-6.1%) of PBL.

Large granular lymphocyte expansions in patients with Felty's syndrome: analysis using anti-T cell receptor V beta-specific monoclonal antibodies.

Felty's syndrome (FS), the association of rheumatoid arthritis (RA) and idiopathic neutropenia, remains an unexplained phenomenon. HLA-DR4 is found in over 90% of cases. Patients with FS may have a T cell lymphocytosis of CD3+CD8+CD57+ large granular lymphocytes (LGL syndrome). In this study of 47 patients with FS, 19% had clear evidence for LGL expansions, while in total 42% had variable evidence for the LGL syndrome using currently available techniques. Of these T cell expansions, 76% were clonal, as demonstrated by Southern blotting and analysis with T cell receptor (TCR) beta chain constant region probes. This technique may fail to detect clonal populations in some patients. Cytofluorographic analysis using antibodies specific for TCR V beta chains identified patients with clonal LGL expansions with results comparable to those obtained with Southern blotting. No evidence for shared V beta usage among expansions from different patients was seen. The role of LGL in RA and FS is currently unclear, but this technique offers a practical and accessible means of identifying patients with LGL expansions, as a starting point for further investigation.

A Rapid and Sensitive Chemiluminescent Immunoassay of Total Thyroxin with DMAE · NHS-Labeled

A novel chemiluminescence immunoassay (CLIA) of total thyroxin (TT4) with the acridinium ester of DMAE · NHS [2′,6′-dimethyl-4′-(n-succinimidyloxycarbonyl) phenyl-10-methyl-acridinium-9-carboxylate methosulfate] labeled has been developed. In our method, microwells were coated with anti-T4 monoclonal antibody (McAb) and DMAE · NHS synthesized by our laboratory was conjugated with streptavidin (SA). T4-BSA was conjugated with biotin-N-hydroxysuccinimide (B · NHS), T4-BSA- B · NHS and the T4 in the standard or sample competitively react with anti-T4 McAb. Streptavidin-biotin separation and enhanced techniques were applied in this assay. The presented approach shows many excellent characteristics, particularly, rapid assay process and high detection sensitivity.

The effect of gene therapy using CTLA4Ig/silica‐nanoparticles on canine experimental autoimmune thyroiditis

The present study aimed to determine the effect of canine CTLA4Ig on canine autoimmune thyroiditis. In a previous study, we established a canine model of autoimmune thyroiditis by immunizing normal dogs with bovine thyroglobulin. An in vitro study using recombinant CTLA4Ig revealed that this protein can inhibit the expression of Th1-type cytokines and the pro-inflammatory cytokines tested. As a result of the in vitro study, we constructed therapeutic CTLA4Ig/silica-nanoparticles and applied them to the treatment of experimentally induced canine autoimmune thyroiditis. Gene therapy resulted in significant reductions in anti-canine-thyroglobulin autoantibody titer, anti-T4 antibody titer and T-cell proliferation against thyroglobulin and in the mRNA expressions of interleukin-18 in fresh peripheral blood mononuclear cells (PBMC) from all dogs. There was also a significant reduction compared to day 0 in tumor necrosis factor-alpha and interferon-gamma levels in the supernatant from cultured PBMC. The CTLA4Ig-induced suppression of Th1 cytokines is relatively more significant than it appears because autoimmune thyroiditis is a Th1-polarized disease. Thus, CTLA4Ig can improve Th1/Th2 cytokine balance in autoimmune thyroiditis by downregulating Th1 cytokines.

DCC and TS protein expression in resected gastric cancer: A Hellenic Cooperative Oncology Group Study

Abstract There is a high risk of relapse after resection of gastric cancer. We studied the prognostic significance of the deleted colorectal cancer (DCC) gene and thymidylate synthase (TS) protein expression after resection of gastric cancer. Protein expression in the primary tumor of 146 patients with serosal and/or lymph node involvement was studied immunohistochemically by using anti-DCC and anti-TS monoclonal antibodies. DCC expression was found in 69.9%, while low TS staining intensity (0+,1+) and focal staining (&lt;25% of tumor cells stained) were found in 44.6% and 33.8%, respectively. Overall survival (OS) was significantly longer in patients with DCC (p=0.014) negative tumors. TS expression was not an independent prognostic factor. Lack of DCC expression was associated with significantly longer cause-specific survival (CSS) (p=0.040) after curative resection. In conclusion, DCC expression is an independent prognostic factor in patients undergoing resection of gastric cancer while TS expression was not associated with the prognosis in our study.