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PubMed1981

Studies with a monoclonal antibody on the distribution of Thy-1 in the lymphoid and extracellular connective tissues of the dog.

BALB/c mice were immunized with canine T lymphocytes from lymph node and used in cell fusion experiments to derive monoclonal antibodies to the T lymphocyte surface. The cloned hybrid line F3-20-7 described in this paper was shown to be secreting antibodies directed at canine Thy-1 since (1) the tissue distribution of the antigen corresponded precisely to the tissue distribution expected of canine Thy-1 from previous studies and (2) the antigen purified from F3-20-7 monoclonal antibody affinity columns (a) had the same mobility on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) as pure rat Thy-1 and (b) could inhibit and assay shown previously to be directed at canine Thy-1 on the basis of cross-reactivity with rat Thy-1. Studies with the fluorescence-activated cell sorter showed that all thymocytes and lymph node T lymphocytes of the dog were Thy-1 positive, establishing that Thy-1 is a T cell marker in the dog, as in the mouse, although the presence of a small percentage of Thy-1-positive B cells could not be entirely excluded. In addition, 15% of the nucleated bone marrow cells were Thy-1 positive, which is similar to the finding in rat bone marrow. Localization studies on frozen sections of skin demonstrated large amounts of Thy-1 organized in a highly structured manner around the basal parts of the hair follicles. This covering of Thy-1 was lost as the follicles approached the surface. Epidermal cells were Thy-1 negative. In the kidney, Thy-1 was shown to be strongly associated with all of the basement membranes of the medulla. The kidney cortex, including glomeruli was essentially negative, except for the occasional staining of basement membranes, possibly of collecting ducts.

Animals↗
PubMed2004

Effective photoimmunotherapy of murine colon carcinoma induced by the combination of photodynamic therapy and dendritic cells.

PURPOSE: The unique mechanism of tumor destruction by photodynamic therapy (PDT), resulting from apoptotic and necrotic killing of tumor cells accompanied by local inflammatory reaction and induction of heat shock proteins (HSPs), prompted us to investigate the antitumor effectiveness of the combination of PDT with administration of immature dendritic cells (DCs). EXPERIMENTAL DESIGN: Confocal microscopy and Western blotting were used to investigate the influence of PDT on the induction of apoptosis and expression of HSP expression in C-26 cells. Confocal microscopy and flow cytometry studies were used to examine phagocytosis of PDT-treated C-26 cells by DCs. Secretion of interleukin (IL)-12 was measured with ELISA. Cytotoxic activity of lymph node cells was evaluated in a standard (51)Cr-release assay. The antitumor effectiveness of PDT in combination with administration of DCs was investigated in in vivo model. RESULTS: PDT treatment resulted in the induction of apoptotic and necrotic cell death and expression of HSP27, HSP60, HSP72/73, HSP90, HO-1, and GRP78 in C-26 cells. Immature DCs cocultured with PDT-treated C-26 cells efficiently engulfed killed tumor cells, acquired functional features of maturation, and produced substantial amounts of IL-12. Inoculation of immature DCs into the PDT-treated tumors resulted in effective homing to regional and peripheral lymph nodes and stimulation of cytotoxic activity of T and natural killer cells. The combination treatment with PDT and administration of DCs produced effective antitumor response. CONCLUSIONS: The feasibility and antitumor effectiveness demonstrated in these studies suggest that treatment protocols involving the administration of immature DCs in combination with PDT may have clinical potential.

Animals↗
PubMed1990

Kinetic immunochemical studies of IgG production during local and systemic anti-influenza immune response in rats.

The kinetics of anti-influenza IgG antibodies in serum and nasal wash during the local and systemic immune response in rats was studied. The influenza virus A/HK/1/68 (H3N2) was injected by two different routes--intranasally and subcutaneously in the hind footpads. The proliferation of the Ig-forming cells in the popliteal and paratracheal lymph nodes either local or distant according to the mode of virus administration was also studied. The results obtained during the primary and secondary immune response suggested that an atypical immunization also produced a strong immune response in the distant lymph nodes. The nature of the secondary immune response supports the concept of migration of the activated lymphocytes from the peripheral lymph nodes to the natural portal of entry of virus thus giving rise to specific clonal population.

Administration, Intranasal↗
PubMed2006

Inhibition of macropinocytosis blocks antigen presentation of type II collagen in vitro and in vivo in HLA-DR1 transgenic mice.

Professional antigen-presenting cells, such as dendritic cells, macrophages and B cells have been implicated in the pathogenesis of rheumatoid arthritis, constituting a possible target for antigen-specific immunotherapy. We addressed the possibility of blocking antigen presentation of the type II collagen (CII)-derived immunodominant arthritogenic epitope CII259-273 to specific CD4 T cells by inhibition of antigen uptake in HLA-DR1-transgenic mice in vitro and in vivo. Electron microscopy, confocal microscopy, subcellular fractionation and antigen presentation assays were used to establish the mechanisms of uptake, intracellular localization and antigen presentation of CII by dendritic cells and macrophages. We show that CII accumulated in membrane fractions of intermediate density corresponding to late endosomes. Treatment of dendritic cells and macrophages with cytochalasin D or amiloride prevented the intracellular appearance of CII and blocked antigen presentation of CII259-273 to HLA-DR1-restricted T cell hybridomas. The data suggest that CII was taken up by dendritic cells and macrophages predominantly via macropinocytosis. Administration of amiloride in vivo prevented activation of CII-specific polyclonal T cells in the draining popliteal lymph nodes. This study suggests that selective targeting of CII internalization in professional antigen-presenting cells prevents activation of autoimmune T cells, constituting a novel therapeutic strategy for the immunotherapy of rheumatoid arthritis.

Amiloride↗
PubMed1991

[The humoral immune response of different strains of mice after intragastrointestinal immunization with human sperm antigens].

The inbred Balb/c and C57 mice, and the outbred Swiss Webster mice were intragastrointestinally immunized with human sperm antigens. The lymphocytes from the spleen, mesenteric lymph node (MLN), Peyer's patch (PP) and uterus or epididymis were isolated and cultured. The lymphocyte-secreting antisperm IgG and IgA and the antisperm antibodies in the gut wash and serum were determined with enzyme-linked immunosorbent assay (ELISA). In the Balb/c and Swiss Webster mice, the immune responses to sperm have shown to be stronger than that in C57, stronger in female than in male. The antigenicity of sperm membrane extracts seems to be higher than that of whole sperm. Antisperm antibodies secreted by lymphocytes from the epididymis and uterus have demonstrated to be detectable. For stimulation of the local immune response, the intra-PP and intralumina immunizations are more effective than others.

Animals↗
PubMed1999

[RT-PCR assay for detecting PSA mRNA in peripheral blood of prostate cancer patients].

A sensitive technique using reverse transcription and polymerase chain reaction (RT-PCR) has been used to detect circulating tumor cells in the peripheral blood of men with prostate cancer. We evaluated the clinical utility of this method for staging and monitoring for prostate cancer. Peripheral blood from 39 patients with prostate cancer and 7 non-prostate cancer controls was analyzed for prostate specific antigen (PSA) messenger RNA (mRNA) using RT-PCR. In 8 among 22 patients (36.4%) with clinically localized prostate cancer (T2 or T3), PSA mRNA was detected by RT-PCR (RT-PCR positive). Five out of 8 patients (62.5%) with regional lymph node and/or bone metastases were RT-PCR positive. The number of patients with RT-PCR positive was more frequent in a higher clinical stage. In 22 patients with clinically localized prostate cancer, 2 of the 9 patients who underwent radical prostatectomy had positive surgical margins and both patients were RT-PCR positive. Six of the 7 patients with negative surgical margin were RT-PCR negative. In the 9 cases that had been treated by combined antiandrogen blockade for metastatic prostate cancer, 4 patients whose serum PSA level were less than 4 ng/ml were all RT-PCR negative. More over 4 of 5 patients with more than 4 ng/ml of serum PSA level were RT-PCR positive. All control samples were RT-PCR negative. This study suggested that this technique using RT-PCR may provide useful information in treating patients with prostate cancer, especially for candidates for radical prostatectomy. The value of this modality as a prognostic factor awaits for further follow-up.

Biomarkers, Tumor↗
PubMed2001

Telomerase activity and expression of human telomerase RNA component and human telomerase reverse transcriptase in lung carcinomas.

The aim of this study was to evaluate the usefulness of determination of telomerase activity and expression of human telomerase RNA component (hTERC) and human telomerase reverse transcriptase (hTERT) for the diagnosis of lung carcinomas. The tissues studied consisted of 115 carcinomas and adjacent nonneoplastic lung, which were removed surgically without previous chemotherapy or radiotherapy. Telomerase activity was determined using a semiquantitative polymerase chain reaction-based telomeric repeat amplification protocol (TRAP) assay. The results obtained were classified into high and low telomerase groups. Localization of expression was examined by using in situ hybridization and immunohistochemistry. The correlation between telomerase activity in lung carcinoma and clinicopathologic features, including prognosis, was investigated. Telomerase activity in lung carcinomas was detected in 107 of 115 (93%) lung carcinomas, but not in any adjacent noncancerous tissues, and was significantly higher in small cell carcinoma than in any other histologic type. This activity also was significantly higher in poorly differentiated than in well-differentiated squamous cell carcinomas and adenocarcinomas. The overall survival rate (P =.020) was significantly lower in the high telomerase group. Messenger RNAs for hTERC and hTERT were mainly detected in the cytoplasm of cancer cells by in situ hybridization, and TERT protein was localized in the nuclei of these cells by immunohistochemical staining. Determinations of telomerase activity by in situ hybridization, immunohistochemistry, and TRAP assay are useful for evaluating the diagnosis and prognosis of lung carcinomas.

Adenocarcinoma↗
PubMed2004

Role of natural killer cells in the rejection process of corneal allografts in rats.

BACKGROUND: The exact mechanism of human corneal allograft rejection, which is the major cause of corneal transplant failure, remains unclear. We investigated the role of natural killer (NK) cells in rat corneal allograft rejection by examining the aqueous humor (AH) cell infiltrate on different postoperative days. METHODS: Flow cytometric analysis was performed on the AH and submandibular draining lymph node (DLN) cells before transplantation and at different time points thereafter. In addition, we performed functional cytotoxicity assays with cells present in the AH during corneal rejection. RESULTS: We demonstrated a gradual increase in the absolute cell number of different hematopoietic subpopulations in the AH after allogeneic cornea transplantation. CD3CD4 cells, mainly monocytes and macrophages, were the predominant subpopulation 2 days after transplantation, followed by a successive relative increase of CD4 T cells, CD8 T cells, CD161 T cells, and NK cells. NK and CD161 T cells were present at a 10- to 15-fold higher percentage than in the DLN, suggestive of local expansion of these cells. A higher percentage of NK cells were CD8-negative compared with DLN NK cells. AH cells specifically lysed allogeneic cells, and this cytotoxicity was mainly attributable to NK cells but not to CD4 or CD8 T lymphocytes. CONCLUSION: These results confirm the crucial role of CD4 cells in the allogeneic corneal graft rejection process and implicate NK cells as possible mediators of the rejection.

Animals↗
PubMed1991

Prostatic specific antigen related to clinical status 1 to 14 years after radical retropubic prostatectomy.

Serum prostatic specific antigen (PSA by Tandem-R immunoassay) was measured in 190 patients after radical prostatectomy for prostate cancer. Serial measurements were made in all patients operated on in the past 3 years; 131 had undetectable levels and 59 had levels in the detectable range. Only 10% of the patients with undetectable PSA following surgery had seminal vesical involvement or positive lymph nodes. None of the patients with undetectable PSA have had clinical recurrence within the 41 months of mean follow-up. In 14 patients who had PSA serially measured since surgery, detectable levels were found within 6 months of operation and 7 of these patients had clinical progression within 2 years; 39 patients had detectable PSA after radical prostatectomy with no clinical evidence of recurrence. Biopsy of the anastomosis was performed in 11 patients with isolated detectable PSA after surgery and local recurrence was established in 4. PSA was detectable later in the follow-up of 45 patients operated on before the PSA assay became available but the date when PSA actually became detectable is not known. A relatively new method of estimating that date and constructing a corresponding Kaplan Meier curve is presented. PSA is an effective marker for monitoring patients after radical prostatectomy as it often detects early persistent disease in patients with detectable measurements 6 months post-operatively or recurrent disease in patients with later rising levels.

Aged↗
PubMed1981

The role of T cell sets in the rejection of a methylcholanthrene-induced sarcoma (S1509a) in syngeneic mice.

The ability of different T cell sets to confer protection in mice against a methylcholanthrene-induced sarcoma, S1509a, was examined. Intravenous infusion of lymph node and spleen cells from A/J donors immunized with S1509a into normal A/J recipients retarded subcutaneous growth of S1509a but did not lead to complete eradication of the tumor during a 9-day period of observation. This protective effect was lost if the transferred cells were treated with anti-Thy 1.2 and complement. The ability of different populations of lymphoid cells to retard tumor growth after inoculation with tumor cells subcutaneously was examined (Winn assay). Nylon-wool-passed cells from lymph nodes and spleens of tumor immunized animals were treated either with anti-Ly 1.2 or with anti-Ly 2.2 antiserums and complement and inoculated with tumor cells in normal A/J mice. The tumor was measured daily for 10 or more days. Ly l cells and unfractionated T cells efficiently suppressed tumor growth; Ly 23 cells had little or no effect. When small numbers of Ly 1 cells were injected along with twice as many Ly 23 cells, the growth of the tumor was also inhibited. Histologic examination of inoculated sites at 24-72 hours after local transfer showed a more intense mononuclear infiltrate in animals inoculated with tumor cells and T cells from immunized animals than in animals given injection with tumor cells alone, or with tumor cells and T cells from nonimmunized animals. The findings indicate that Ly 1 cells are capable of retarding the growth of the sarcoma, presumably by eliciting a delayed hypersensitivity reaction. By contrast, Ly 23 cells, which can mediate cytotoxicity, had little or no effect on tumor growth.

Animals↗
PubMed1988

Intraoperative radioimmunolocalization of colorectal carcinoma with a hand-held gamma probe and MAb B72.3: comparison of in vivo gamma probe counts with in vitro MAb radiolocalization.

A new intraoperative approach to tumor localization using radiolabelled monoclonal antibody (MAb) B72.3 involves the use of a hand-held gamma-detecting probe (GDP) by the surgeon and, subsequently, the pathologist. We report here the use of 125I-labelled MAb B72.3 IgG and a GDP to localize primary and metastatic colorectal cancer in 31 patients. The patients were administered radiolabelled MAb i.v., and all underwent surgical exploration 5 to 35 days post-injection. In vivo localization of the MAb was evaluated using a GDP, with tumor and normal tissue counts being obtained. In each case, the subsequent tumor and normal tissue that were resected were analyzed in vitro for MAb localization; this was evaluated by calculating the radiolocalization index, i.e., the ratio of the injected dose per gram localized to tumor versus that of normal tissue. When the GDP was used intraoperatively, MAb B72.3 localized tumors in 68% (21/31) of the patients; the arbitrary criterion of tumor-to-normal tissue ratios higher than or equal to 2.0:1 in vivo being taken as positive. Resected tumor radiolocalization indices ranged from 0.5 to 543, and 71% (22/31) of the patients studied had tumors with radiolocalization indices higher than or equal to 3. Of 50 carcinoma biopsies, 34 that were probe-positive were antigen-positive when B72.3 was used in immunoperoxidase assays, while 4 carcinoma biopsies that were probe-negative were also antigen-negative. Twelve of 50 biopsies were probe-negative and antigen-positive, but many of these lesions only contained a few antigen-positive cells; none of the 50 was probe-positive and antigen-negative. Tumors of all histologic grades localized injected MAb and, in general, higher in vivo probe ratios and radiolocalization indices were obtained from patients who underwent surgery 20 to 35 days following injection of the MAb. MAb B72.3 localized tumor in all sites to which colon carcinoma commonly metastasizes, including mesenteric and peri-aortic lymph nodes, liver, lung, and peri-rectal soft tissue. There was a strong statistical correlation (p = 0.001) between detecting MAb B72.3 localization to tumors using the GDP intraoperatively and subsequent in vitro analysis of cpm/g for tumor versus normal tissues. These studies thus further validate the use of 125I-labelled MAb B72.3 IgG and of a hand-held gamma probe for the intraoperative detection of carcinomatous lesions.

Adult↗
PubMed1986

Activated T cells with immunoregulatory functions at different sites of involvement in sarcoidosis. Phenotypic and functional evaluations.

Cells from recovered BAL fluid and from infiltrates in different involved tissues (lungs, lymph nodes, conjunctiva, liver, spleen, and skin) were studied in 22 patients with active sarcoidosis in order to define the surface phenotype, functional in vitro properties, and topographic distribution of the cells in granulomatous lesions. Our data demonstrated a compartmentalization of activated T cells with immunoregulatory functions from the blood to all sites of disease activity. In fact, these cells were found to express the T4+ Leu 8- 5/9+ T17- phenotype, which belongs to cells with helper activity, and that provide heightened responses in functional assays of helper activity, IL-2 release, and the ability to respond in AMLR's. Both a cellular redistribution and a local in vitro replication account for this tissue compartmentalization in sarcoidosis. The microanatomic location of activated T cells, as defined by immunohistological evaluation, showed that the state of activation in these T cells may be a consequence of an intimate contact between helper cells and macrophages within the sarcoidosis granulomas.

Adult↗
PubMed1996

CD4 expression is important but not essential for infection with exogenous mouse mammary tumor virus.

We studied local events in the popliteal lymph nodes of CD4-deficient mice following foot pad injection with an MMTV strain which carries the gene for a V beta 14-specific superantigen. Injection of the V beta 14-specific MMTV induced vigorous expansion of V beta 14+ CD4+ T cells and B cells in their lymph nodes of CD4+/- heterozygous control mice. On the other hand, CD4-/- mice injected with the MMTV showed a proliferation of V beta 14+ T cells among the population of TCR alpha beta + CD4-CD8- T cells, although to a lesser extent. This phenomenon was not accompanied by vigorous B cell expansion. A PCR assay revelated that the MMTV definitely infected the lymph nodes cells of the CD4-/- mouse. However, the infectivity of the MMTV in CD4-/- mice was approximately 20 times lower than that in CD4+/- mice. These findings indicate that, in MMTV infection of CD4-deficient mice, the superantigen-reactive T cells among the population of TCR alpha beta +CD4-CD8- T cells substitute for the superantigen-reactive CD4- T cells of normal mice, and that the absence of CD4 molecules decreased the infectivity of MMTV because of insufficient expansion of the superantigen-reactive T cells.

Animals↗
PubMed2002

Identification of Corynebacterium pseudotuberculosis isolates from sheep and goats by PCR.

The present study was carried out to estimate the prevalence of caseous lymphadenitis (CL) in sheep and goats slaughtered at the local abattoir in Elazig province located in the east of Turkey, between September and December 2000. A total of 2046 sheep and 2262 goat carcasses were examined during the study period and 118 abscessed lymph nodes, 89 from sheep and 29 from goats, were collected. Corynebacterium spp. strains were isolated from 81.4% of the abscesses, giving an overall prevalence of 2.2%. The prevalence was 3.5 and 1.1% in sheep and goats, respectively. PCR on DNA extracted from 96 suspicious isolates, using a pair of Corynebacterium pseudotuberculosis-specific primers, was positive for 93. Although cross-reaction with C. ulcerans, a human/bovine species, was observed, the PCR assay used in this study may successfully be applied for the diagnosis of CL in goats and sheep as an alternative to conventional methods, owing to its advantages of specificity and speed.

Animals↗
PubMed2004

Mode of spread in the early phase of lymphatic metastasis in pancreatic ductal adenocarcinoma: prognostic significance of nodal microinvolvement.

BACKGROUND: The aim of this study was to assess the prognostic significance of nodal microinvolvement as well as the mode of spread in the early phase of lymphatic metastasis in patients with node-negative pancreatic ductal adenocarcinoma. METHODS: Lymph nodes from 48 node-negative patients with R0 resected pancreatic ductal adenocarcinoma were sampled from 3 different compartments: 1) distal hepatoduodenal ligament, 2) superior-anterior compartment, and 3) posterior-inferior. Tissue sections of 148 lymph nodes classified as tumor free by routine histopathology were examined, using a sensitive immunohistochemical assay with the antiepithelial monoclonal antibody Ber-EP4 for tumor cell detection. With regard to histopathologic tumor staging and grading, 26 (54.2%) of the patients were staged as pT1/pT2, 22 (45.8%) as pT3/pT4, while 31 (64.6%) as G1/G2 and 17 (35.4%) patients as G3. Of the 148 "tumor free" lymph nodes, 56 contained Ber-EP4-positive tumor cells. These 56 lymph nodes were from 28 of the 48 patients. The multivariate Cox regression analysis revealed the independent prognostic impact of nodal microinvolvement on relapse-free and overall survival. Analysis by compartment, from which the lymph nodes were collected, revealed that overall survival time (P = 0.006) and time to local recurrence (P = 0.015) depend on the presence of nodal microinvolvement in the superior-anterior compartment. CONCLUSIONS: The influence of occult tumor cell dissemination in lymph nodes of patients with histologically proven pancreatic ductal adenocarcinoma supports the need for further tumor staging through immunohistochemistry. This could be a helpful tool in proper selection of patients for adjuvant chemotherapy.

Biomarkers, Tumor↗
PubMed1979

Preferential homing of passively transferred T cells into skin allografts of mice.

An assay which uses two differentially labeled cell populations was used to characterize the preferential localization of passively transferred syngeneic cells immunized to specific alloantigens. Splenocytes cytotoxic to B10.D2 and B10.BR alloantigens were harvested from (C57BL/6 X A/J)F1 (B6AF1) donors bearing acutely rejected skin allografts. One population was labeled in vitro with 3H-thymidine and the other with 14C-thymidine. The labeled cells were pooled and then transferred i.v. into B6AF1 hosts bearing 5-day-old skin grafts from B10.D2 and B10.BR donors. After 48 hr the mice were killed, and the relative amount of cells present in the skin grafts and draining axillary lymph nodes was derived by comparing the 3H:14C ratios of the harvested tissues. The results of these studies indicated that cytotoxic splenocytes harvested from donors bearing acutely rejected skin allografts preferentially localize to the relevant skin allograft after passive systemic transfer. The homing behavior of these splenocytes was augmented by T cell enrichment and significantly diminished by pretreatment with anti-Thy-1.2 serum plus rabbit complement. There was no evidence of preferential homing within the draining axillary lymph nodes. It can be concluded that a T cell population derived from in vivo sensitized splenocytes exhibits preferential homing to relevant skin allografts upon passive transfer.

Animals↗
PubMed2005

Induction of a proinflammatory program in normal human thyrocytes by the RET/PTC1 oncogene.

Rearrangements of the RET receptor tyrosine kinase gene generating RET/PTC oncogenes are specific to papillary thyroid carcinoma (PTC), the most frequent thyroid tumor. Here, we show that the RET/PTC1 oncogene, when exogenously expressed in primary normal human thyrocytes, induces the expression of a large set of genes involved in inflammation and tumor invasion, including those encoding chemokines (CCL2, CCL20, CXCL8, and CXCL12), chemokine receptors (CXCR4), cytokines (IL1B, CSF-1, GM-CSF, and G-CSF), matrix-degrading enzymes (metalloproteases and urokinase-type plasminogen activator and its receptor), and adhesion molecules (L-selectin). This effect is strictly dependent on the presence of the RET/PTC1 Tyr-451 (corresponding to RET Tyr-1062 multidocking site). Selected relevant genes (CCL20, CCL2, CXCL8, CXCR4, L-selectin, GM-CSF, IL1B, MMP9, UPA, and SPP1/OPN) were found up-regulated also in clinical samples of PTC, particularly those characterized by RET/PTC activation, local extrathyroid spread, and lymph node metastases, when compared with normal thyroid tissue or follicular thyroid carcinoma. These results, demonstrating that the RET/PTC1 oncogene activates a proinflammatory program, provide a direct link between a transforming human oncogene, inflammation, and malignant behavior.

Blotting, Western↗
PubMed2002

Preoperative blood reverse transcriptase-PCR assays for prostate-specific antigen and human glandular kallikrein for prediction of prostate cancer progression after radical prostatectomy.

The association of the results reverse transcription-PCR (RT-PCR) assays for prostate-specific antigen (PSA) with prostate cancer stage and progression remains controversial. We have previously shown that RT-PCR amplification of a native human glandular kallikrein fragment (hK2-L), but not a splice-variant hK2 transcript (hK2-U), performed preoperatively on peripheral blood before radical prostatectomy is an independent predictor of metastases to lymph nodes [K. M. Slawin, et al., Cancer Res., 60: 7142-7148, 2000]. We evaluated the ability of preoperative peripheral blood RT-PCR for PSA and hK2 to predict prostate cancer progression after surgery. The study group consisted of 224 consecutive patients who underwent radical prostatectomy for clinically localized disease (median follow-up: 52.9 months). Also evaluated were 14 healthy men and 8 patients with hormone-naïve, prostate cancer metastatic to bone. RT-PCR for PSA, hK2-L, and hK2-U were positive in 24, 25, and 26%, respectively, of prostatectomy patients; 88, 71, and 86%, respectively, of patients with metastases to bones; 7, 14, and 36%, respectively, of healthy men. Preoperative RT-PCR-hK2-L positivity was associated with higher final Gleason sum (P = 0.028), with an increased risk of metastases to lymph nodes (P = 0.019) and with overall and aggressive disease progression (P = 0.0231 and P = 0.028, respectively). Preoperative blood RT-PCR-PSA and RT-PCR-hK2-U were not associated with characteristics or outcomes of prostate cancer. Preoperative blood RT-PCR for PSA and hK2-U are neither therapy-guiding staging tools nor prognostic indicators in patients with clinically localized prostate cancer. In contrast, preoperative RT-PCR-hK2-L status is associated with established markers of aggressive prostate cancer.

Adult↗