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Enhancement of therapeutic effects of recombinant interleukin 2 on a transplantable rat fibrosarcoma by the use of a sustained release vehicle, pluronic gel.

We have tested the feasibility of pluronic F-127 gel (PLF-127; a polyoxyethylene-polyoxypropylene surface active block copolymer) as a sustained release vehicle for topical administration of interleukin 2 (IL-2) in order to enhance the therapeutic effects of IL-2 against a rat fibrosarcoma, KMT-17. Injection of human DNA recombinant IL-2 (3 X 10(4) units s.c.) in 30% (w/w) PLF-127 into rats provided detectable serum IL-2 levels for up to 10 h, while injection of IL-2 alone provided detectable IL-2 levels for 3 h. When, following s.c. inoculation with 1 X 10(5) KMT-17 tumor cells into rats, IL-2 (6 X 10(4) units/day) in PLF-127 gels was injected s.c. around the growing tumor inoculum every 2 days for 10 days from Day 1 to Day 19, the survival days of rats were more prolonged [mean survival day, 32.3 +/- 5.4 (SD)] as compared with that of rats treated with saline [20.7 +/- 2.1] than mean survival days of rats treated with IL-2 alone [27.3 +/- 4.5] or PLF-127 alone [22.9 +/- 3.3]. Moreover, the span of mean survival days of rats treated with IL-2 in PLF-127 locally (31.7 +/- 5.9) was much longer than that of rats given IL-2 in PLF-127 systemically (22.8 +/- 3.4). By means of a Winn assay, stronger tumor neutralizing activities were observed in regional lymph node cells obtained from tumor bearing rats treated with IL-2 in PLF-127 than were observed in lymph node cells from rats treated with IL-2 alone or PLF-127 alone (percentage of inhibition, 90.3, 12.2, and -15.5%, respectively). The therapeutic effects of IL-2 were thus found to be consistent with the antitumor activity in regional lymph node cells. These results suggest that the enhanced therapeutic effects of IL-2 in PLF-127 are due to enhancement of antitumor immune responses induced by sustained IL-2 activity at the tumor sites.

Animals↗

The relationship between storage and secretion of specific antibody by immune lymphoid cells: ultrastructural localization of anti-peroxidase antibodies in plaque-forming cells of the rabbit popliteal lymph node.

The ultrastructure of antibody-forming cells (AFC) has been studied in the lymph node cell population from rabbits locally immunized with horseradish peroxydase (PO) incorporated in complete Freund's adjuvant, and the kinetics of AFC development followed from day 7 to day 18 after one injection of PO. Identification of the AFC was done by local hemolysis assay, using carboxymethyl cellulose solidifying medium and PO-coated sheep erythrocytes. AFC were thereafter transferred by micromanipulation into a Beem capsule, fixed, treated by PO for fine ultrastructure detection of anti-PO antibodies, included, sectioned and studied by electron microscopy. It was found that the AFC were essentially of three categories: lymphocytes, proplasmacytes and plasmacytes, with (+), or without (-), intracellular antibody. The proportion of these categories varies with the time elapsed since the injection of antigen and with the plaque-forming activity of the population: lymphocytes (-) are relatively more numerous (over 30%) at the early stages of immunization (day 7). The number of plasmacytes increases with immunization. If most of them contain intracellular antibody at the early stages (up to day 9), the proportion of plasmacytes (+) decreases markedly afterwards. The cell type distribution is compatible with the idea that the lymphocytes are the precursors of plasmacytes, proplasmacytes being transitional forms, but no direct filiation scheme can actually be deduced from these experiments.

Animals↗

Reverse transcriptase polymerase chain reaction for prostate specific antigen in the management of prostate cancer.

PURPOSE: Polymerase chain reaction is a powerful tool for expanding minute quantities of deoxyribonucleic acid (DNA) for detailed study. Reverse transcriptase polymerase chain reaction (RT-PCR) involves the initial conversion of messenger ribonucleic acid (mRNA) to DNA, followed by the amplification of the DNA product for molecular analysis. The mRNA for prostate specific antigen (PSA) is expressed only by prostatic cells. RT-PCR offers a potentially more sensitive assay for the detection of cells expressing PSA mRNA in peripheral circulation or in extraprostatic tissues. The current status of RT-PCR in the amplification and detection of PSA gene expression in the management of prostate cancer is reviewed. MATERIALS AND METHODS: The literature was reviewed for available data using RT-PCR for detection of prostatic cells outside of the prostate. RESULTS: Amplification of mRNA by RT-PCR represents a highly sensitive method of detection of gene expression. A single cell expressing PSA among 10 to 100 million lymphocytes can be detected by the RT-PCR assay. This assay may detect extraprostatic or circulating prostatic cells in peripheral blood, lymph nodes and bone marrow in many patients with prostate cancer. Various studies have reported sensitivities of detection of PSA expressing cells in the peripheral blood ranging from 0 to 88%. This wide range in the sensitivity may be partly due to tremendous variation between the protocols used in each study. In patients with lymph node metastasis the RT-PCR assay appears more reliable than immunohistochemistry for identification of prostatic tissue in the lymph node. In some series analyses of radical prostatectomy specimens suggest a strong correlation between a positive RT-PCR result and capsular invasion by the tumor, while others do not support its use in determining pathological stage. In the majority of reports the RT-PCR assay was highly specific for detection of extraprostatic PSA expression in prostate cancer patients, and negative for detection in men with benign prostatic hyperplasia and in women. Sources of potential false-positive and false-negative results of this assay are identified and discussed. CONCLUSIONS: RT-PCR can detect PSA expressing cells that are otherwise undetectable by other means in patients with localized and metastatic cancers. This assay is highly specific, since PSA expressing cells were consistently undetectable in the peripheral circulation of patients without prostate cancer in most studies. Limited data to date suggest that this test may have a role in the staging of tumors before radical prostatectomy and in the serial followup of patients after treatment. RT-PCR may improve the detectability of lymph node metastasis over immunohistochemistry. Presently this test should remain a powerful research tool in the study of the biology and behavior of prostate cancer, and it should not be used to guide any clinical decision making. The use of this assay as a prognostic and management tool for prostate cancer is in the earliest stages.

Clinical Trials as Topic↗

High endothelial venule binding as a predictor of the dissemination of passaged murine lymphomas.

It has long been postulated that normal lymphocyte homing mechanisms help determine the metastatic spread of lymphoid neoplasms. The traffic of normal lymphocytes is controlled in part by the regulated expression of surface receptors for high endothelial venules (HEV), specialized venules that mediate the extravasation of circulating lymphocytes from the blood into lymphoid organs and sites of chronic inflammation. Here we have compared the in vivo growth patterns of HEV-binding vs. nonbinding murine lymphomas passaged intramuscularly into syngeneic recipients. We report that lymphomas that bind well to HEV (as assessed in a quantitative in vitro assay) disseminate widely via the blood, involving all lymph node groups symmetrically. Although both HEV-binding and nonbinding lymphomas gain access to the blood, gross involvement of lymph nodes by nonbinding lymphomas is limited to nodes draining local tumor at the site of injection, a prominent feature of these lymphomas; distant lymph nodes are not enlarged. The results suggest that the expression of functional receptors for HEV either controls the hematogenous dissemination of malignant lymphocyte populations to HEV-bearing organs, or is coregulated with factors determining this metastatic behavior. The findings support the concept that normal lymphocyte homing mechanisms are important to the spread of leukemias and lymphomas.

Animals↗

Peptide-specific T cell response to Mycobacterium tuberculosis: clinical spectrum, compartmentalization, and effect of chemotherapy.

The T cell repertoire of 59 patients with untreated tuberculosis was compared with that of 46 bacille Calmette-Guérin-vaccinated controls by assaying the proliferative responses to six permissively recognized peptides from the 16-, 19-, and 38-kDa molecules of Mycobacterium tuberculosis. A trend from higher to lower reactivity following this order: vaccinated controls > lymph node disease > localized extrapulmonary > pulmonary > pleural was seen for 4 of the peptides (P < .03). The decreased response of blood lymphocytes from patients with pleural tuberculosis was partially accounted for by sequestration of peptide-responsive cells within the pleural fluid. Chemotherapy "reversed" the depressed proliferative responses of patients with pulmonary and pleural tuberculosis depending on the peptide origin, being greatest for peptides of 16 kDa, transient for those of 19 kDa, and least for those of 38 kDa. These data demonstrate antigen specificity in the decreased responsiveness of patients with tuberculosis.

Adolescent↗

Hematogenous spread of malignant melanoma cells in different stages of disease.

Patients with malignant melanoma and distant metastases generally have an unfavorable prognosis, with a median survival of about 6 months. The mechanisms of hematogenous spread and implantation of melanoma cells are, however, poorly understood, because the standard diagnostic methods are not sensitive enough to detect oligocellular micrometastases. Recently a method using reverse transcription and polymerase chain reaction to determine tyrosinase mRNA in peripheral blood, which indicates the presence of circulating melanoma cells, has been developed. We utilized this assay to examine blood samples of 56 patients with malignant melanoma in different stages of disease. In one of 10 patients in stage I (localized disease) and in six of 17 patients with regional lymph nodes metastases (stage II) tyrosinase mRNA was detected. Tyrosinase transcripts were found in all 29 patients with distant metastases (stage III). Interestingly, tyrosinase mRNA was also detected in six patients with metastatic amelanotic malignant melanoma. In contrast, tyrosinase mRNA was not detectable in any of 39 healthy subjects or 17 patients with other malignancies. These findings may be helpful to define a patient group at high risk for systemic spread of disease.

Base Sequence↗

Comparison of local and systemic responsiveness of lymphocytes in vitro to Bovicola ovis antigen and concanavalin A in B. ovis-infested and naive lambs.

The in vitro proliferation assay was used to determine lymphocyte responsiveness to soluble antigen of B. ovis and to Concanavalin A (Con A) in peripheral blood, spleen and various lymph nodes from B. ovis-infested and naive lambs. From March to July, an assay of monthly blood samples showed generally higher proliferative responses to antigen and Con A in B. ovis-infested than naive lambs. The proliferative response of cells from the skin-draining prescapular lymph nodes to B. ovis antigen was significantly higher in B. ovis-infested than naive lambs. Responses of cells from the medial iliac, mediastinal and mesenteric lymph nodes (which do not receive lymph from the skin) and spleen showed no significant differences between groups. Within the B. ovis-infested lambs, the response of cells from the prescapular lymph node was significantly higher than that from any other lymphoid organ examined. Responsiveness of the prescapular, medial iliac and mesenteric lymph node and spleen cells to Con A was not significantly different between groups, while mediastinal lymph node cells showed a significantly higher response in B. ovis-infested lambs. The data indicate that the antigen-specific cellular immune response is operating mainly locally, at the level of the skin and draining lymph nodes. Responses to the T cell mitogen Con A did not support non-specific immunodepression as reported in other ectoparasite/host systems.

Animals↗

Should we reconsider the indications for ileo-obturator node dissection with localized prostate cancer?

OBJECTIVE: To evaluate the need for ileo-obturator node dissection in patients with localized prostate cancer who are undergoing radical retropubic prostatectomy. PATIENTS AND METHODS: Over a 5-year-period, 95 patients underwent bilateral pelvic lymphadenectomy. Ninety were performed in association with planned radical prostatectomy and five were staging procedures in clinical stage T3 patients prior to radiotherapy or hormonal therapy. The patients with localized prostate cancer were stage T1a (one patient), T1b (21), T2a (30), and T2b (38). Pre-operative biopsies in the patients with localized cancer were well differentiated in 44 patients, moderately well differentiated in 45 and poorly differentiated in one. In the patients with T3 tumours, pre-operative biopsies were well differentiated in one, moderately well differentiated in two and poorly differentiated in two. Prostate-specific antigen (PSA) levels ranged from 0.4 to 110.1 ng/mL (Hybritech assay). RESULTS: Two patients had positive lymph node dissections on fixed section. These two patients had well-differentiated T1b disease with a PSA level of 72.4 ng/mL and poorly differentiated T3 disease with a PSA level of 58.5 ng/mL. There was significant upstaging (P < 0.001) and upgrading (P < 0.001) on pathological examination. None of the 71 patients with a PSA < or = 10 ng/mL had positive lymph nodes compared with 8.3% of the 24 patients with a PSA > 10 ng/mL (P = 0.0618). Lymph node metastases were present in 1% of patients with well or moderately well-differentiated prostate cancer on pre-operative biopsy versus 33% with poorly differentiated disease (P = 0.0625). These P values strongly suggest an association, achieving significance only at the 10% level which might be the more appropriate level given the relative lack of power of the study due to the small number of patients with positive lymph nodes. CONCLUSION: These results suggest that routine ileo-obturator node dissection in patients with well or moderately well-differentiated, localized prostate cancer and a PSA level < 10 ng/mL may be unnecessary, especially as a separate procedure.

Aged↗

Clinical benefit of chemosensitivity test for patients with regional lymph node-positive esophageal squamous cell carcinoma.

BACKGROUND AND OBJECTIVES: Patients who have undergone resection for lymph node-positive esophageal squamous cell carcinoma are at high risk of recurrence and early death. The role of the postoperative adjuvant chemotherapy in this population needs to be determined. The present study was designed to determine whether the chemosensitivity test in fresh human esophageal squamous cell carcinoma, using highly purified tumor cells, correlates with the clinical response. METHODS: A retrospective review of all patients with resected squamous cell carcinoma of the thoracic esophagus between 1993 and 2000 was performed. We determined the chemosensitivity for cisplatin (CDDP), 5-fluorouracil (5-FU), mitomycin C, and adriamycin in vitro in fresh human esophageal squamous cell carcinoma using the MTT assay. Regional lymph node-positive (N1) patients who received sequential postoperative chemotherapy were compared with lymph node positive patients who underwent surgery alone. RESULTS: A total of 50 patients were reviewed, and chemosensitivity tests were successfully performed in 46 patients: 20 patients received surgery alone (S group), and 26 patients received surgery plus postoperative chemotherapy (SC group) according to results of MTT assay using highly purified tumor cells. When the SC group was divided into an SC-low group (inhibition rate of CDDP + 5-FU was below 85%, n = 15) and an SC-high group (over 85%, n = 11), the SC-high group showed more significant survival prolongation than the S group or the SC-low group (P < 0.01). CONCLUSIONS: Our results suggest that the results of the conventional MTT assay may be useful in evaluating the optimum adjuvant chemotherapy for patients with regional lymph node positive (pN1) esophageal squamous cell carcinoma.

Aged↗

Whole body positron emission tomography/computed tomography staging of metastatic choroidal melanoma.

PURPOSE: To evaluate whole-body positron emission tomography (PET)/computed tomography in staging of patients with metastatic choroidal melanoma. DESIGN: Interventional non-randomized clinical study. METHODS: Twenty patients were referred for whole-body 18-fluoro-2-deoxy-D-glucose (FDG) PET/computed tomography imaging because of suspected metastatic choroidal melanoma. PET/computed tomography images were studied for the presence and distribution of metastatic melanoma. Subsequent biopsies were performed to confirm the presence of metastatic disease. RESULTS: Twenty patients underwent PET/computed tomography. Eighteen were imaged because of abnormal clinical, hematologic, or radiographic screening studies during the course of their follow-up after plaque brachytherapy or enucleation. Two were imaged before treatment of their primary tumor. PET/computed tomography revealed or confirmed metastatic melanoma in eight (40%) of these 20 patients. The mean time from initial diagnosis to metastasis was 47 months (range 0 to 154). The most common sites for metastases were the liver (100%), bone (50%), lung (25%), lymph nodes (25%), and subcutaneous tissue (25%). Cardiac, brain, thyroid, and posterior abdominal wall lesions (12.5%) were also noted. Six patients (75%) had multiple organ involvement. No false positives were noted. PET/computed tomography imaging also detected benign lesions of the bone and lymph nodes in three patients (15%). All patients had hepatic metastases and liver enzyme assays were abnormal in only one (12.5%) of eight patients. CONCLUSIONS: PET/computed tomography imaging is a sensitive tool for the detection and localization of hepatic and extra-hepatic (particularly osseous) metastatic choroidal melanoma.

Aged↗

Phase I dose escalation clinical trial of adenovirus vector carrying osteocalcin promoter-driven herpes simplex virus thymidine kinase in localized and metastatic hormone-refractory prostate cancer.

Osteocalcin (OC), a major noncollagenous bone matrix protein, is expressed prevalently in prostate cancer epithelial cells, adjacent fibromuscular stromal cells, and osteoblasts in locally recurrent prostate cancer and prostate cancer bone metastasis [Matsubara, S., Wada, Y., Gardner, T.A., Egawa, M., Park, M.S., Hsieh, C.L., Zhau, H.E., Kao, C., Kamidono, S., Gillenwater, J.Y., and Chung, L.W. (2001). Cancer Res. 61, 6012-6019]. We constructed an adenovirus vector carrying osteocalcin promoter-driven herpes simplex virus thymidine kinase (Ad-OC-hsv-TK) to cotarget prostate cancer cells and their surrounding stromal cells. A phase I dose escalation clinical trial of the intralesional administration of Ad-OC-hsv-TK followed by oral valacyclovir was conducted at the University of Virginia (Charlottesville, VA) in 11 men with localized recurrent and metastatic hormone-refractory prostate cancer (2 local recurrent, 5 osseous metastasis, and 4 lymph node metastasis) in order to determine the usefulness of this vector for the palliation of androgen-independent prostate cancer metastasis. This is the first clinical trial in which therapeutic adenoviruses are injected directly into prostate cancer lymph node and bone metastasis. Results show that (1). all patients tolerated this therapy with no serious adverse events; (2). local cell death was observed in treated lesions in seven patients (63.6%) as assessed by TUNEL assay, and histomorphological change (mediation of fibrosis) was detected in all posttreated specimens; (3). one patient showed stabilization of the treated lesion for 317 days with no alternative therapy. Of the two patients who complained of tumor-associated symptoms before the treatment, one patient with bone pain had resolution of pain, although significant remission of treated lesions was not observed by image examination; (4). CD8-positive T cells were predominant compared with CD4-positive T cells, B cells (L26 positive), and natural killer cells (CD56 positive) in posttreated tissue specimens; (5). levels of HSV TK gene transduction correlated well with coxsackie-adenovirus receptor expression but less well with the titers of adenovirus injected; and (6). intrinsic OC expression and the efficiency of HSV TK gene transduction affected the levels of HSV TK protein expression in clinical specimens. Our data suggest that this form of gene therapy requires further development for the treatment of androgen-independent prostate cancer metastasis although histopathological and immunohistochemical evidence of apoptosis was observed in the specimens treated. Further studies including the development of viral delivery will enhance the efficacy of Ad-OC-hsv-TK.

Adenoviridae↗

Vaginal transmission of HIV-1 in hu-SCID mice: a new model for the evaluation of vaginal microbicides.

OBJECTIVE: To develop an animal model of vaginal transmission of HIV-1 for the evaluation of vaginal microbicides. DESIGN: Vaginal infection was performed in SCID mice reconstituted with 4 x 107 human peripheral blood lymphocytes (hu-PBL) by non-invasive vaginal administration. The hu-PBL were previously infected in vitro with a non-syncytium (NSI) strain of HIV-1 (SF162) (hu-PBL-SCID). Lymphocyte migration in vivo was examined using fluorescently labelled human lymphocytes. METHODS: The percentage of CD4 T cells, plasma viral load and p24 antigen were evaluated using fluorescent activated cell sorting (FACS), the Amplicor HIV-1 monitor kit and enzyme-linked immunosorbent assay, respectively. Polymerase chain reaction (PCR) analysis was performed on DNA extracted from spleen and lymph nodes. For in vivo migration of labelled lymphocytes, the mice were sacrificed after 4, 24 and 48 h; vaginae and local lymph nodes were removed, snap frozen with OCT, sectioned and examined by fluorescent microscopy and FACS. RESULTS: HIV transmission was established using virus-infected cells inoculated vaginally, as shown by FACS, HIV viral load, p24 and PCR results. Labelled cells were easily located within the vaginal tissues after 4 h. However, few or no cells could be identified after 24 or 48 h at the vaginal level, whereas labelled cells could be detected at the level of regional lymph nodes. CONCLUSIONS: Because of its simplicity and practical features compared with other animal models, the vaginal HIV-infected hu-SCID mouse model may prove useful to test the activity of compounds against cell-associated HIV and, possibly, other sexually transmitted diseases.

Animals↗

Follow-up with serum Cyfra 21-1 in patients with squamous cell carcinomas of the head and neck.

OBJECTIVE: Finding tumor markers for disease progression, and especially development of distant metastases, is desirable for patients with squamous cell carcinoma of the head and neck (SCCHN). Elevated serum levels of Cyfra 21-1 (cytokeratin fraction 21-1) have been frequently associated with disease progression in patients with lung cancer. In SCCHN, Cyfra 21-1 has not been established as a routine tumor marker yet, probably due to difficulties in finding the appropriate cut-off for the serum level. The aim of this study was to investigate whether assessment of changes in serum Cyfra 21-1 over time can predict distant metastases in patients with SCCHN, without attempting to establish an arbitrary cut-off for abnormal levels. METHODS: Cyfra 21-1 serum levels of 25 patients with SCCHN and distant metastases were evaluated by means of an ELISA test kit. RESULTS: There was a wide range of Cyfra 21-1 serum levels at the time of primary diagnosis, without correlation with tumor size, lymph node status, time to recurrence, or presence of distant metastases. All patients had a clear increase of Cyfra 21-1 levels which preceded the appearance of distant metastases clinically. CONCLUSIONS: Due to the wide range of Cyfra 21-1 levels at the time of primary tumor diagnosis, Cyfra-21-1 is neither a suitable screening marker for SCCHN, nor for diagnosis of distant metastases at the time of initial diagnosis of the tumor, but is of evident prognostic value for follow-up, especially for early detection of distant metastases.

Antigens, Neoplasm↗

[Treatment of colorectal cancer].

The large majority of colorectal cancers are well or moderately differentiated adenocarcinomas. Their biological behavior is not as malignant as that of stomach cancer, with a tendency of slow growth and limited spread. Surgery is therefore the first choice for management of these cancers even in cases where complete removal of the tumor is not expected, surgery is useful for the prolongation or improvement of the quality of life. However, the correct choice of operation for such cases is very important. The extent of resection and dissection must be decided based upon the degree of cancer spread. Both must be sufficient, but not excessive. In cases in the early stage where cancerous growth has not spread beyond the submucosal layer, local or segmental excision may be a good enough treatment. On the other hand, for far advanced rectal cancer involving adjacent organs, combined resection and pelvic evisceration is curatively effective. Several pathological findings of resected specimens influencing prognosis were investigated in the present study. In cases which appeared to be diffuse infiltrative spreading type in macroscopic appearance or poorly differentiated, undifferentiated, or mucinous types of cancer in their histological features, survival rates were low. However, these were uncommon and over half of them were much too far advanced to receive curative resection. Irradiation combined with hyperthermia or heavy chemotherapy were applied, but were mostly ineffective. For these uncommon types of cases as well as far advanced cases, we have no effective treatment other than surgery at this time.

Adenocarcinoma↗

Partial purification of a common antigen in bovine lymphoma and its use in a lymphocyte blastogenesis assay.

An antigen was isolated from tumor cells derived from cases of bovine lymphoma which caused the in vitro blastogenesis of lymphocytes from nine of 13 (69%) adult cattle with lymphoma. Lymphocytes derived from five of 122 (4%) normal adult cattle also underwent blastogenesis while lymphocytes from three cases of the sporadic form of lymphoma did not respond. Blastogenesis of lymphocytes from normal cattle may have resulted from alloantigen activity in the antigen; however, normal lymph node antigen did not stimulate lymphocytes from normal adult cows or cows with lymphoma. Antigen derived from tumor was localized to the cell membrane and cytoplasm of tumor cells by indirect immunofluorescent staining using an antiserum prepared in rabbits. Specific fluorescence was reduced by the absorption of this antiserum with a crude tumor extract. The antiserum did not cross-react with normal lymph node antigen or viral components as demonstrated by immunodiffusion. With further refinement of the antigen, the blastogenesis assay may be of use in the early detection and study of the pathogenesis of lymphoma in adult cattle.

Animals↗

Differences in the migration of B and T lymphocytes: organ-selective localization in vivo and the role of lymphocyte-endothelial cell recognition.

The migration of B and T lymphocytes in the mouse has been studied by using 1) short-term in vivo homing studies, and 2) an in vitro assay of lymphocyte binding to specialized lymphoid organ venules (post-capillary, high endothelial venules (HEV)) in frozen sections of lymph nodes and Peyer's patches. The homing characteristics of B and T cell populations are largely independent of their organ of origin. B cells from any source distribute preferentially to Peyer's patches, whereas T cells home preferentially to peripheral lymph nodes. This organ specificity of migration appears to be determined at the site of lymphocyte exit from the blood by selective recognition of organ-specific determinants on the endothelial cells of HEV. In addition, the in vivo tendency of B cells to migrate preferentially to the spleen, and of T cells to localize better in lymph nodes is confirmed. The results indicate that, in a hypothetical situation in which an equal number of B and T lymphocytes localized in peripheral lymph nodes (or bound in vitro to peripheral node HEV), there would be about 2.5 B cells for every T cell in the mesenteric node, four to six B cells per T cell in Peyer's patches, and seven to nine B cells per T cell in the spleen. Comparison of these homing preferences with the distribution of B and T lymphocyte populations in situ suggests that selective lymphocyte migration may help determine the proportions of functionally distinct lymphocyte classes in particular lymphoid organs or sites of chronic inflammation, and thus may serve to influence the character of local immune responses.

Animals↗

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↗

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↗