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Results for “Local Lymph Node Assay”

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PubMed1996

Multiple tumour marker assays in advanced cervical cancer: relationship to chemotherapy response and clinical outcome.

Serum levels of squamous cell carcinoma antigen (SCC), CA 125 and CA 15.3 were measured in 102 patients with locally advanced cervical cancer undergoing neoadjuvant chemotherapy (NACT) and radical surgery. We found a significant correlation between SCC concentration and stage, histotype, cervical tumour size and lymph node status. For CA 125 and CA 15.3, no significant difference in the distribution of marker levels according to histopathological variables was found. In a multivariate analysis, histological type, FIGO stage and SCC positivity (> 5 ng/ml) proved to be independent predictors of response to neoadjuvant chemotherapy. Moreover, logistic regression analysis showed that CA 15.3 may be a significant adjunct to SCC in the prediction of chemotherapy response. Of the three markers tested, only CA 125 was significantly related to patient survival. In the multivariate analysis, clinical response to chemotherapy and CA 125 status (> 35 U/ml) retained an independent prognostic value. Our data suggest that the tumour markers used in this study could be useful in the management of locally advanced cervical cancer. Pretreatment serum levels of SCC, together with CA 15.3 assay, may be a useful tool in the determination of response to chemotherapy, while CA 125 assay could be evaluated as a prognostic risk factor in these patients.

Adult↗
PubMed2001

Isolation of a novel human lung-specific gene, LUNX, a potential molecular marker for detection of micrometastasis in non-small-cell lung cancer.

We have isolated a novel human lung-specific gene, LUNX (lung-specific X protein), by differential-display mRNA analysis. The full-length cDNA contained 1,015 nucleotides including an open reading frame of 768 nucleotides encoding 256 amino acids. We localized the gene to chromosomal region 20p11.1-q12 by radiation hybrid mapping. Using an RT-PCR assay specific for LUNX mRNA, 35 non-small-cell lung-cancer (NSCLC) tumors and 0 of 16 normal lymph nodes were positive. Furthermore, LUNX mRNA expression was enhanced in 26 (84%) of 31 NSCLC tumors vs. corresponding cancer-free lung tissues by semi-quantitative analyses with multiplex RT-PCR. We assessed the possibility of LUNX mRNA as a molecular marker for detection of micrometastasis in dissected lymph nodes obtained from 20 patients with NSCLC tumors. LUNX mRNA was detected in 16 (80%) of 20 histologically positive lymph nodes and 21 (25%) of 84 histologically negative lymph nodes. Comparative analyses of the conventional histological examination and the RT-PCR detection assay for LUNX mRNA showed that the detection rate of metastases in lymph nodes by the RT-PCR assay was higher in 12 and consistent in 6 of the total 20 NSCLC patients. We demonstrate that the LUNX RT-PCR assay is a potential diagnostic method for detection of micrometastases in lymph nodes of NSCLC patients.

Amino Acid Sequence↗
PubMed1981

Age-dependent allospecific change in susceptibility to local lymph node GVHR in rats.

Inoculation of parental strain lymphocytes into the footpads of young F1 hybrid rats induces GVHR, which can be quantitated by estimation of the weight of the popliteal lymph node 7 days later. In a number of strain combinations adult and old F1 animals were also useful for the assay. In some, however, it was observed that F1 rats did not support GVHR to the same extent as young did, when one of the parental strains served as donor for the GVHR, but reacted normally if the other parental strain was used. This phenomenon was analysed in serum and cell transfer experiments as well as in vitro in MLC. It was possible to transfer suppression in vivo, whereas some degree of specific MLC inhibition was observed when serum from old F1 animals undergoing GVHR was assessed. The 'old' pattern of response could be reverted partially by splenectomy and completely by low dose irradiation of the recipient before GVHR.

Aging↗
PubMed1988

Definition of the pulmonary antibody response to ovalbumin following local challenge in systemically immunized rats.

The in vivo pulmonary immune response of rats to local stimulation with antigen was assessed by measuring antigen-specific antibody and antibody-secreting cells utilizing enzyme-immunoassay technology. Sprague-Dawley rats were immunized subcutaneously with ovalbumin (OA) emulsified in Freund's incomplete adjuvant, challenged with OA intratracheally on Day 19 and sacrificed 1, 2, 3, or 4 days later. Specific antibody-secreting cells in the lung-associated lymph nodes were enumerated with the ELISA-SPOT assay and antibody concentration in the pulmonary lavage fluids and sera was assessed with the ELISA. The greatest response for each parameter was on Day 2. Cellular infiltration of the lung was minimal. Cellular infiltrates consisted mainly of polymorphonuclear leukocytes and were most numerous in the lavage fluid on Days 1 and 2 and in the lung parenchyma on Day 2 after challenge. Local production versus serum transudation of antibody was evaluated by comparing the levels of OA-specific antibody to albumin in the lavage fluid and serum. The data showed that antibody in the lungs was primarily produced locally.

Animals↗
PubMed2002

High-sensitive 2nd generation thyroglobulin immunoradiometric assay. Clinical application in differentiated thyroid cancer management.

BACKGROUND: Circulating human thyroglobulin (hTG) measurements have a pivotal role in the management of patients affected by differentiated thyroid cancer (DTC). The present study was undertaken by employing a new developed high-sensitive hTG immunoradiometric assay to evaluate its diagnostic performance in patients affected by radically cured and relapsing DTC and to set the most appropriate cut-off point for DTC management. METHODS: We retrospectively selected 172 patients without signs of recurrence after primary treatment and 45 patients with recurrences from DTC. Serum samples were collected during l-thyroxine (T4) suppressive therapy (onT4) and 4 weeks after T4 withdrawal (offT4) and hTG measured by a specific high-sensitive IRMA assay (DYNOtest Tg-plus, BRAHMS Diagnostica GmbH, Berlin, Germany). Sera showing the presence of AbhTG or hTG-recovery less than 80% were excluded from the study. ROC curve analysis was performed to select the best cut-off levels and diagnostic performance of the marker evaluated. RESULTS: By using onT4 cut-off level of 0.2 ng/mL and offT4 cut-off level of 0.5 ng/mL we obtained a sensitivity/specificity/accuracy profile of 0.91/0.98/0.96 and 0.98/0.97/0.97, respectively. We found onT4-hTG false-negative results in 4 patient with local recurrence (n=2) or cervical lymph-node metastasis (n=2) while only 1 patient with local recurrence showed negative offT4-hTG. However, onT4 and offT4-hTG false-negative results were observed in 9 and 5 patients when 1.0 ng/mL cut-off level was employed. CONCLUSIONS: On the basis of our data, we conclude that DYNOtest Tg-plus assay is very effective and accurate in the evaluation of patients with DTC.

Adolescent↗
PubMed2005

XAGE-1 mRNA expression in prostate cancer and antibody response in patients.

To evaluate the feasibility of cancer vaccine targeting XAGE-1, we investigated the expression of 4 XAGE-1 transcript variants and the humoral immune response to XAGE-1 in prostate cancer patients. XAGE-1a, b, c, d mRNA expression was analyzed in 54 prostate cancer specimens and 8 specimens of benign prostate hypertrophy (BPH) by reverse transcription-polymerase chain reaction (RT-PCR). The humoral response to XAGE-1 was investigated in sera obtained from 278 patients with prostate cancer and 40 healthy volunteers by enzyme-linked immunosorbent assay (ELISA) using recombinant protein. XAGE-1b mRNA expression was observed in 14 of 54 (26%) prostate cancer specimens, while XAGE-1a, c, and d mRNA expressions were observed in 1, 1, and 3, respectively. None of the 4 XAGE-1 transcript variants was observed in the 8 BPH specimens. Antibody against XAGE-1 was detected in sera from 2 of 129 stage D2 patients, whereas none of sera from 149 patients with localized prostate cancer or lymph node metastasis had detectable XAGE-1 antibody. No reactivity to XAGE-1 was found in sera from the 40 healthy individuals.

Aged↗
PubMed1990

Prostate specific antigen and local recurrence after radical prostatectomy.

We evaluated the location of recurrent disease in 63 patients with carcinoma of the prostate who had abnormal levels of prostate specific antigen (greater than 0.4 ng./ml., Tandem-R assay) 6 to 240 months after radical prostatectomy but who were otherwise considered to be without evidence of disease. The evaluation involved physical examination including digital rectal examinations by 3 urologists, isotopic bone scans, computerized tomography scans of the abdomen and pelvis, cystoscopy, and random needle biopsies of the urethrovesical anastomotic area. In 6 patients metastatic disease to the bone and/or lymph nodes was found and local prostate cancer was discovered in 5. Among 57 patients without evidence of disease by the usual methods of evaluation needle biopsies of the anastomosis revealed local disease in 42%. No local disease was discovered in 30 post-radical prostatectomy patients with normal prostate specific antigen levels. There was a wide range of transrectally palpable contours after radical prostatectomy in patients with and without elevated prostate specific antigen levels. We conclude that prostate specific antigen but not digital rectal examination is an excellent early indicator of possible local recurrence after radical prostatectomy. Whether the prevalence of local disease after radical prostatectomy using prostate specific antigen levels and needle biopsies of the anastomosis is greater than heretofore has been appreciated will require further study.

Aged↗
PubMed1995

Identification of an HLA-A11-restricted epitope from the tandem repeat domain of the epithelial tumor antigen mucin.

Epithelial cell mucin encoded by the MUC-1 gene is overexpressed and aberrantly glycosylated on pancreatic, breast, and ovarian cancers as well as on multiple myelomas. It is recognized by patients' Ab and by T cells derived from tumor-draining lymph nodes. The T cell recognition is not MHC restricted and is specific for an epitope previously localized to the immunodominant tandem repeat region of the native mucin molecule. In search of possible MHC-restricted epitopes in the same immunodominant region, we synthesized a panel of overlapping, nine-amino acid long peptides spanning the MUC-1 tandem repeat and first examined their binding to specific human MHC class I molecules using two independent flow cytometry-based assay systems. This approach identified one peptide, p9-17 (STAPPAHGV), that bound to HLA-A1, -A2.1, -A3, and -A11. Measurements of the affinity of binding to each of these alleles, using a quantitative molecular binding assay, indicated that only the relative binding affinity to HLA-A11 was close to immunogenic values. We tested the immunogenicity of p9-17 in vitro. We detected a secondary T cell response specific for p9-17 in lymph nodes from an HLA-A11 breast cancer patient. Moreover, CTL specific for p9-17 peptide could be generated from PBL in several healthy HLA-A11 donors by primary in vitro stimulation.

Amino Acid Sequence↗
PubMed2004

Culture and serologic survey for Mycobacterium avium subsp. paratuberculosis infection among southeastern white-tailed deer (Odocoileus virginianus).

From July 1998 through October 2002, radiometric culture (ileocecal lymph node, mesenteric lymph node, and feces) and serologic testing by enzyme-linked immunosorbent assay (ELISA) were used to survey white-tailed deer (Odocoilens virgianus) from the soutlheastern United States for infection by Mycobacterium avium subsp. paratuberculosis (Mptb), the causative agent of paratuberculosis (Johne's disease). Mycobacterium avium subsp. paratuberculosis was isolated from the ileocecal lymph node of one of 313 deer (0.3%) originating from 63 populations in Alabama, Arkansas, Florida, Georgia, Kentucky, Louisiana, Maryland, Mississippi, North Carolina, South Carolina, Tennessee, and West Virginia (USA). Six deer (2%), all from different populations, had ELISA results above a 0.25 sample-to-positive cutoff value, but none of the ELISA reactors originated from the population from which the single Mptb isolation was made. These six deer were seronegative when tested by agar gel immunodiffusion (AGID). Collectively, these data indicate that white-tailed deer currently do not constitute a broad regional reservoir for Mptb; however, further study is warranted to clarify the significance, if any, of infected deer to the epizootiology of paratuberculosis on a local scale. Adaptation and validation of an ELISA or another serologic assay for use with deer and other wildlife would markedly enhance Mptb surveillanece among wild populations and would be a powerful tool for gaining information on the role of wild species in epidemiology of paratuberculosis.

Animals↗
PubMed2006

Pre-clinical study of 213Bi labeled PAI2 for the control of micrometastatic pancreatic cancer.

PURPOSE: The urokinase plasminogen activator (uPA) and its receptor (uPAR) are expressed by pancreatic cancer cells and can be targeted by the plasminogen activator inhibitor type 2 (PAI2). We have labeled PAI2 with (213)Bi to form the alpha conjugate (AC), and have studied its in vitro cytotoxicity and in vivo efficacy. METHODS AND MATERIALS: The expression of uPA/uPAR on pancreatic cell lines, human pancreatic cancer tissues, lymph node metastases, and mouse xenografts were detected by immunohistochemistry, confocal microscopy, and flow cytometry. Cytotoxicity was assessed by the MTS and TUNEL assay. At 2 days post-cancer cell subcutaneous inoculation, mice were injected with AC by local or systemic injection. RESULTS: uPA/uPAR is strongly expressed on pancreatic cancer cell lines and cancer tissues. The AC can target and kill cancer cells in vitro in a concentration-dependent fashion. Some 90% of TUNEL positive cells were found after incubation with 1.2 MBq/ml of AC. A single local injection of approximately 222 MBq/kg 2 days post-cell inoculation can completely inhibit tumor growth over 12 weeks, and an intraperitoneal injection of 111 MBq/kg causes significant tumor growth delay. CONCLUSIONS: (213)Bi-PAI2 can specifically target pancreatic cancer cells in vitro and inhibit tumor growth in vivo. (213)Bi-PAI2 may be a useful agent for the treatment of post-surgical pancreatic cancer patients with minimum residual disease.

Animals↗
PubMed1996

Preoperative lymph-node staining with liposomes containing patent blue violet. A clinical case report.

A case study is reported using patent blue violet entrapped in liposomes to localize lymph nodes before surgery. Liposomes containing 44 mg patent blue violet were injected into foot lymphatics of a patient due to undergo retroperitoneal staging-lymphonodectomy. Lymph nodes were readily visualized and removed, and urine and serum was collected for assay of the dye. The maximum concentration of patent blue violet in serum was 2.7 micrograms mL-1, 1.5 h after the termination of the injection, almost no dye was detectable by the time of the operation 24 h later. In the first urine sample (2 h after the termination of the injection) the concentration of patent blue violet was 1.3 micrograms mL-1, and subsequently diminished, with the last urine sample to contain detectable dye being collected 20 h later. A total of 0.8 mg dye (2% of the injected amount) was excreted in the urine suggesting 10% of the circulating amount of dye was renally excreted. Endolymphatic injection of patent blue violet simplifies the often difficult process of finding the lymph nodes and additionally reduces side-effects. All surgery cases demanding a radical lymphonodectomy may profit, including testicular or bladder cancers and many gynaecological tumours.

Adult↗
PubMed1978

Importance of controlling the regional draining lymph nodes in breast cancer.

A major issue in the treatment of breast cancer deals with local control of disease and the necessity for removing microscopically involved lymph nodes. Many radiation oncologists are looking to the parasternal lymphatics as possible renewed justification for postoperative radiation. It is concluded that prospectively designed studies are needed to assay the importance of the internal mammary nodes and their adequate control. Lymphoscintigraphy may offer a method of evaluating these nodes without radical surgery.

Axilla↗
PubMed2000

Nondissection of pelvic lymph nodes does not influence the results of perineal radical prostatectomy in selected patients.

OBJECTIVES: Retrospective studies have shown that pelvic lymph node dissection can be dispensed with in selected men undergoing radical prostatectomy. We prospectively evaluated the influence of nondissection of pelvic lymph nodes on tumor progression in our first 100 perineal prostatectomies. METHODS: From October 1992 to February 1998, 100 patients underwent radical perineal prostatectomy for localized prostate cancer. Preoperative PSA, the Gleason score of positive biopsies and age at surgery were noted. Forty-three of the 100 patients (group 1) did not undergo pelvic lymph node dissection because their preoperative PSA level was below 10 ng/ml (Hybritech assay, normal value 4 ng/ml) and the Gleason score of their positive biopsies was below 7. These 43 patients were compared with 25 of the 114 patients operated on during the same period by the retropubic approach and who had pelvic node dissection and the same preoperative criteria (PSA <10 ng/ml and a Gleason score of positive biopsies <7; group 2). All prostatectomy specimens were processed according to the Stanford protocol: prostate weight, Gleason score, capsular, seminal vesicle, lymph node and surgical margin status, and tumor volume were studied. Postoperative followup was based on routine serum PSA assays after 1 and 3 months and then half-yearly. Biological progression was defined as PSA level which was detectable postoperatively (>/=0.2 ng/ml). Kaplan-Meier analysis was used to evaluate the likelihood of biochemical recurrence. Results were compared by using Fisher's test, the Mann-Whitney test and the log-rank test. Differences were considered significant when the p value was <0.05. RESULTS: No differences in preoperative characteristics were observed; in groups 1 and 2, mean age was 65.9 and 64.7 years, PSA was 6.7 and 5.11 ng/ml, and the Gleason biopsy score was 5.7 vs. 5.0, respectively. In groups 1 and 2, specimen weight was 44.5 and 54.3 g (p = 0.04), the Gleason score was 6.2 and 5.6, tumor volume was 0.91 and 0.8 ml, 81.4 and 84% of patients were in stage pT2, 13.9 and 12% had extracapsular disease, 4.6 and 0% had seminal vesicle invasion, and 13.9 and 16% had positive surgical margins, respectively. The mean follow-up was 2.74 years (0.27-5.59 years). The actuarial 5-year recurrence-free rate was 78% in group 1 and 80% in group 2 (p>0.05). CONCLUSION: The lack of pelvic lymph node dissection does not influence the intermediate term results of perineal radical prostatectomy in selected patients (preoperative PSA <10 ng/ml and Gleason score for positive biopsies <7).

Aged↗
PubMed2002

A comparison of local and peripheral parasite-specific antibody production in different strains of mice infected with Trichuris muris.

The serum parasite-specific antibody responses of different mouse strains infected with Trichuris muris reflect the nature of the T-helper response mounted by the host, in that resistant Th2-responding strains, such as BALB/K, produce immunoglobulin (Ig)G1 and susceptible predominantly Th1-responding strains, such as AKR, produce IgG2a and IgG1. However, the kinetics of antibody production in the sera, as determined by enzyme-linked immunosorbent assay, do not reflect infection status in that resistant strains can expel their worm burdens before antibodies are detectable in the sera. Here, we show that parasite-specific antibody production by in vitro lipopolysaccharide-stimulated mesenteric lymph node cells (MLN) not only correlate with serum antibody isotypes, but also follow expulsion kinetics. Additionally, the antibody levels seen locally match changes in absolute B220+ cell numbers in the MLN (determined by flow cytometry) and changes in MLN parasite-specific plasma cells in the MLN (determined by ELISPOT). These results show that B cell responses are tightly regulated locally in both resistant and susceptible strains of mice infected with T. muris.

Animals↗
PubMed1986

Production and characterization of a monoclonal antibody to human type III collagen.

Human type III collagen from placenta was isolated and purified for use as an immunogen. A monoclonal antibody was produced which specifically recognizes epitopes unique to type III collagen. The specificity of the antibody was determined by inhibition ELISA, an immunoblot assay, and by immunoprecipitation. Results indicated that the monoclonal antibody recognized only the alpha 1(III) polypeptide chains and did not crossreact with type I, IV, or V collagen. The monoclonal antibody was also used for immunohistochemical localization of type III collagen in tissue sections of human placenta, bovine spleen, and lymph node. In placenta, both large and small blood vessels showed pronounced staining of the tunica media, which contains largely smooth muscle cells, known to synthesize type III collagen. In contrast, the intimal areas and endothelial cells showed no staining with the antibody. In the placental villi, staining was limited to the villous core, where fine fibrillar structures showed strong staining. In lymph nodes, the capsule and pericapsular adipose cells were surrounded by a covering of type III collagen. Within the parenchyma of the node, staining was localized to a branching, reticular array of fine fibers. In the spleen, staining was pronounced in the capsule, splenic trabeculae, and white pulp, where blood vessel staining was especially prominent. The red pulp and splenic sinuses contain little or no type III collagen. The fine network-like or reticular staining pattern found in the lymph node parenchyma is consistent with the staining pattern of the protein reticulin, and suggests that type III collagen may be closely associated with reticulin in certain tissues. Since the role of type III in tissues is unclear, this reagent will be useful in providing new information in this regard.

Amino Acids↗
PubMedDate not supplied

The augmenting effect of OK432-stimulated B cells on the in vitro generation of anti-tumor cytotoxic T lymphocytes from tumor-draining lymph node cells: the possible role of interleukin-12.

Effect of a local injection with a streptococcal preparation OK432 on the in vitro generation of anti-tumor cytotoxic T lymphocytes (CTLs) from tumor-draining lymph nodes (LN) was investigated. A peritumoral injection with OK432 on days 2, 4, 6 and 8 significantly increased both the total cell number and the proportion of B cells in the draining LN cells on day 10 after a subcutaneous inoculation with B16 melanoma. In an in vitro proliferative assay, OK432 showed a stimulatory effect on both normal splenic T and B cells. In a cytolytic assay, the OK432-injected B16-draining LN cells showed a higher level of anti-B16 CTL activity than the B16-draining LN cells after in vitro restimulation. This augmenting effect of OK432 was dependent on the B cells. Moreover, nonadherent cells from the OK432-injected B16-draining LN cells showed a low but significantly higher level of anti-B16 CTL activity than those from the B 16-draining LN cells after in vitro restimulation, whereas this augmenting effect of OK432 was abolished by the in vitro addition of anti-interleukin (IL)-12 monoclonal antibody. Collectively, these findings suggest that the augmenting effect of a local injection with OK432 on the potential of tumor-draining LN cells to turn into anti-tumor CTLs after in vitro restimulation was at least in part due to IL-12 derived from the OK432-stimulated B cells.

Animals↗
PubMed1981

Immune response in the bovine mammary gland after intestinal, local, and systemic immunization.

The immune response in mammary glands of cattle was measured after intestinal, local, and systemic immunization with T4 bacteriophage. Nonlactating pregnant cows were immunized by infusions into the intestine or mammary gland and by subcutaneous injections in the region of the prescapular or external inguinal lymph nodes. Titers of antibodies of different isotypes were measured in serum and in lacteal secretions by enzyme-linked immunosorbent assay, and numbers of cells producing antibodies of each isotype were determined in lacteal secretions by the Jerne plaque assay. Substantial increases in immunoglobulin G subclass 1 (IgG1) and IgG2 antibody titers were detected in serum and lacteal secretions of animals immunized through an intestinal fistula. IgM and IgA antibody responses were low or undetectable. Low numbers of IgA and IgG1 plaque-forming cells were occasionally detected. It is proposed on the basis of these data that migration of antigen-stimulated IgG lymphoblasts, and perhaps of antigen, to spleen and peripheral lymph nodes may be dominant events after intestinal immunization of ruminants. This is consistent with the predominance of serum-derived IgG antibodies in colostrum and milk. Intramammary infusion of antigen gave rise to increases in antibody titers in all classes which were greater not only in lacteal secretions but also in blood serum than with either systemic route used. There was clear evidence from relative antibody titers for local synthesis of antibodies, principally IgA and IgG1, in the immunized glands. Comparison of IgA titers in secretions from the immunized glands with those in serum also suggested that locally synthesized IgA antibodies may have contributed in some measure to serum titers. Local synthesis in both immunized and nonimmunized glands was also reflected by the presence of increased numbers of IgA and IgG1 plaque-forming cells. It is hypothesized that antibody-forming cells responsible for local synthesis originated in lymphoid tissue within the mammary gland or from peripheral lymph nodes, depending upon the route of immunization.

Animals↗
PubMed2006

Cystatin C as a potential marker for relapse in patients with non-Hodgkin B-cell lymphoma.

The concentration of cysteine protease inhibitor cystatin C was determined in sera from 59 patients with non-Hodgkin B-cell lymphoma using ELISA. The sera from 43 age and sex matched healthy blood donors served as controls. Cystatin C was significantly increased in sera of patients without therapy (mean 1136+/-SE 105.7ng/ml, p=0.00001) and with therapy (mean 1073+/-52ng/ml, p=0.001) compared to controls (mean 819+/-28ng/ml). The highest levels were determined in sera of patients with a relapse (mean 1680+/-196ng/ml). By using immunofluorescence staining and confocal microscopy we determined immature dendritic cells as a major population of cystatin C positive cells in affected lymph nodes. Our study reports for the first time that cystatin C is a potential marker for relapse in patients with non-Hodgkin B-cell lymphoma.

Adolescent↗