Clinical and molecular features of adenovirus type 2, 3, and 7 infections in children in an outbreak in Taiwan, 2011.

OBJECTIVES: We studied paediatric patients with human adenovirus (HAdV) infection during the 2011 outbreak in northern Taiwan to define the clinical features of different HAdV genotypes in children. METHODS: Between January and December 2011, 637 patients <19 years of age exhibited culture-confirmed adenoviral infection in Chang Gung Memorial Hospital, and provided specimens available for genotyping by multiplex real-time PCR. Clinical data were collected retrospectively. RESULTS: Excluding five cases with multiple genotypes, 632 cases were included for analysis. Three genotypes were identified, including HAdV-3 (429/632; 67.6%), HAdV-7 (144/632; 22.6%) and HAdV-2 (59/632; 9.8%). Median age was 4.58 years (range 2 months to 18 years), with children infected with HAdV-3 significantly older (82.9% >3 years; p <0.001). Of the 621 inpatients, 98.2% had fevers and all exhibited respiratory symptoms, 75 patients (12.1%) had lower respiratory tract infections, 20 (3.2%) required intensive care (HAdV-2: 1; HAdV-3: 8; and HAdV-7: 11), and three died (all HAdV-7-infected). HAdV-3-infected patients were significantly more likely to have upper respiratory symptoms and a high serum C-reactive protein level >100 mg/L, whereas leucocytosis (white blood cell count >15 000/mm3) was more common in HAdV-2-infected patients (p 0.007). HAdV-7 infections were significantly associated with a longer duration of fever, leucopenia (white blood cell count <5000/mm3), thrombocytopenia (platelet count <150 000/mm3), lower respiratory tract infections, a longer length of hospital stay, and requiring intensive care (all p <0.001). CONCLUSION: Childhood HAdV-2, HAdV-3 and HAdV-7 infections may exhibit different clinical manifestations. Although HAdV-3 was the most prevalent genotype observed during the 2011 Taiwan outbreak, HAdV-7 caused more severe disease characteristics and outcomes.

Hospital-based surveillance and molecular epidemiology of rotavirus infection in Taiwan, 2005-2007.

To determine the distribution of rotavirus strains and facilitate vaccine policy decisions in Taiwan, active hospital-based gastroenteritis surveillance was conducted in three sentinel hospitals. From 1 January 2005 to 31 December 2007, a total of 3435 children less than 5 years old with gastroenteritis were enrolled. The presence of rotavirus was documented by enzyme immunoassay (EIA), and the G and P genotypes were determined by reverse transcription-polymerase chain reaction (RT-PCR) and sequencing methods. Results confirmed that 856 (25%) of these gastroenteritis admissions were EIA-positive for rotavirus and 448 (52%) of the rotavirus positive admissions were less than 2 years old. The most prevalent rotavirus genotypes were G1P[8] (40%), followed by strains G3P[8] (27%), and G9P[8] (17%). These data will help inform decisions as to whether rotavirus vaccine should be considered for inclusion into Taiwan's National Immunisation Programme.

Use of oseltamivir during an outbreak of influenza A in a long-term care facility in Taiwan.

We studied the effectiveness of oseltamivir during an outbreak of influenza A among previously vaccinated patients and staff in a long-term care facility. Seven of 14 staff members and 14 of 41 residents developed either influenza-like illness (ILI) or other respiratory symptoms during a 14-day period from late January to 8 February 2004. On 9 February, therapeutic oseltamivir (75 mg twice daily for five days) was administered to one staff member and seven residents who had developed ILI within the previous 48 h (treatment group). Prophylactic oseltamivir (75 mg once daily for seven days) was administered to 12 staff members and 30 residents who were asymptomatic or whose respiratory symptoms did not meet the diagnosis of ILI (prophylaxis group). The remaining four residents and one staff member had had ILI for more than two days (with subsiding symptoms) and did not receive oseltamivir ('no-oseltamivir' group). None of the 42 subjects in the prophylaxis group developed ILI. Presence of influenza A virus was demonstrated in 24 subjects: seven out of eight in the treatment group, 12 of 42 in the prophylaxis group and all five in the no-oseltamivir group. For confirmation of diagnosis, real-time reverse transcription-polymerase chain reaction was more sensitive than antigen detection and virus isolation. In-time therapeutic and prophylactic oseltamivir successfully interrupted an outbreak of influenza A in a long-term care facility.

Mosaic genome structure of echovirus type 30 that circulated in Taiwan in 2001.

An echovirus type 30 (E30) outbreak occurred in Taiwan in 2001. In this study, one 1998 and nineteen 2001 enterovirus isolates from cerebrospinal fluid (CSF) of children with meningitis were genetically analyzed. Although negative results were obtained using the E30-specific monoclocal antibody in an immunofluorescent assay (IFA) test of all 20 isolates, molecular typing by partial VP1 sequences and subsequent neutralization test identified them as E30. Among those, seven of them were misidentified as echovirus type 4 (E4) when E4-specific monoclonal antibody was used. Complete genome sequences of one E30 isolate (TW-2513) that were IFA-positive to E4 and another (TW-3182) that was IFA-negative to both E30 and E4 were determined and analyzed. The overall percentage nucleotide identity in the structural coding region (P1) between these two isolates is 98.4, while those in the nonstructural regions P2 and P3 are only 83.2 and 84.4, respectively, indicating that the two 2001 Taiwanese E30 strains were probably recombinant. Recombination analysis of these two E30 genomes revealed that their genome structures are mosaic, which might have been formed gradually and frequently over time.

Fcgamma receptor IIa, IIIa, and IIIb polymorphisms of systemic lupus erythematosus in Taiwan.

OBJECTIVE: To determine whether the distribution of Fcgamma receptor IIa, IIIa, and IIIb polymorphisms confers a risk factor for disease susceptibility, and correlates with the clinical characteristics and serological parameters of patients with SLE in Taiwan. METHODS: Genotyping of Fcgamma receptors IIa H/R131, IIIa F/V158, and IIIb NA1/NA2 was performed in 302 patients with SLE and 311 healthy blood donor controls. The distribution of Fcgamma receptor IIa, IIIa, and IIIb genotypes in patients and controls was analysed. Frequencies of three Fcgamma receptor polymorphisms were also compared between lupus patients with and without different clinical manifestations and autoantibodies. RESULTS: No significant skewing in the distribution of Fcgamma RIIa H/R131, Fcgamma RIIIa F/V158, and Fcgamma RIIIb NA1/NA2 was found between patients and controls in Taiwan. The following clinical associations were found: Fcgamma RIIIb NA1/NA1 protected against neuropsychiatric lupus (p = 0.028) but conferred susceptibility to discoid rash (p<0.005); increased Fcgamma RIIIa V/V158 was associated with infections (p = 0.039); increased Fcgamma RIIa H/H131 was associated with earlier onset of lupus (p = 0.01). CONCLUSION: Fcgamma receptor IIa, IIIa, and IIIb polymorphisms may be responsible for the development of distinct manifestations of lupus patients in Taiwan, but there is no significantly skewed distribution in the susceptibility to lupus as a whole.

Levels of interferon-gamma and interleukin-2 receptor-alpha for bronchoalveolar lavage fluid and serum were correlated with clinical grade and treatment of pulmonary tuberculosis.

OBJECTIVE: We investigated possible correlations for interferon-gamma (IFN-gamma) and soluble interleukin-2 receptor-alpha (sIL-2R-alpha) levels in bronchoalveolar lavage fluid (BALF), and clinical grade of pulmonary tuberculosis (TB), which is determined by factors such as extent of pulmonary involvement, fever and loss of body weight. DESIGN: In order to explore these correlations and address associated questions, BALF was collected from 45 patients presenting with active pulmonary TB and 14 healthy controls. Repetitive BALF was collected in 17 patients after 3 months of anti-tuberculosis chemotherapy. The epithelial lining fluid (ELF) levels for IFN-gamma and sIL-2R-alpha were measured using enzyme-linked immunosorbent assay (ELISA) after standardization with urea. RESULTS: Patients with higher-grade pulmonary TB (i.e., with more advanced pulmonary involvement, fever or body weight loss), revealed significantly higher ELF levels for IFN-gamma and sIL-2R-alpha compared to those with lower grade pulmonary TB. Similar results were also determined for sIL-2R-alpha serum levels, but not for IFN-gamma serum levels. After anti-tuberculosis chemotherapy the elevated cytokine levels for ELF and serum significantly decreased in accordance with radiographic improvement. CONCLUSIONS: ELF levels of IFN-gamma and sIL-2R-alpha were correlated with disease grading of pulmonary TB and decreased after anti-tuberculosis chemotherapy.

Development of an ELISA for the detection of serum chromogranin A (CgA) in prostate and non-neuroendocrine carcinomas.

INTRODUCTION: Chromogranin A (CgA) is a glycoprotein found in neuroendocrine cells and may be useful as a tumor marker for neuroendocrine tumors. METHODS: We developed an enzyme-linked immunosorbent assay (ELISA) for serum CgA on a microtiter plate. RESULTS: We established a reference range for both women and men of different age groups ranging from 20 to 80 years. Men appeared to have a slightly higher serum CgA concentration than women. This slight increase in serum CgA concentration was also found in both gender groups with advancing age. We also detected increased serum CgA in a variety of cancers and non-endocrine carcinomas: the majority of the increased serum CgA was associated with specimens containing highly increased concentration of tumor markers. In other words, increased serum CgA was found at later, more advanced stages of the disease in these patients. For patients with prostate cancer, serum CgA was increased much earlier than serum PSA in approximately one-third of prostate cancer patients developing resistance to hormonal therapy. CONCLUSIONS: The early rise of serum CgA provides an early signal for prostate cancer patients who developed resistance to hormonal therapy: this advance signal could create a critical window for therapy changes to be made before diseases progress to a fatal stage.

Amantadine-resistant influenza A virus in Taiwan.

BACKGROUND AND PURPOSE: Amantadine and rimantadine have been used for treatment and prophylaxis of influenza A virus infection. We examined the amantadine susceptibility of field isolates of influenza A virus in Taiwan from 1996 to 1998 to monitor the presence of resistant strains. METHODS: Eighty-four field isolates of influenza A virus were examined for resistance to amantadine by plaque inhibition assay. Virus isolates with amantadine 50% inhibitory concentrations (IC50) greater than 0.9 microgram were chosen for sequence analysis of the M gene that is the molecular target for amantadine/rimantadine. Reverse transcription-polymerase chain reaction (RT-PCR) was used to amplify the viral RNA. RT-PCR products were examined and purified by agarose gel electrophoresis for further sequence analysis. The Genetics Computer Group Sequence Analysis Package and the neighbor-joining method listed in the Molecular Evolutionary Genetic Analysis package were used for phylogenetic analysis. RESULTS: One field strain was amantadine resistant (IC50 > 10 micrograms/mL), with a mutation (position 31, serine to asparagine) in the M2 protein. The resistant virus was isolated from a non-immunocompromised child without a history of amantadine/rimantadine treatment. None of the family members reported previous exposure to amantadine/rimantadine. CONCLUSIONS: In this series, amantadine-resistant influenza A (H1N1) virus was isolated from a non-immunocompromised Taiwanese child without a known history of exposure to this drug. Resistant field isolates were rare. Due to the increasing use of amantadine/rimantadine in Taiwan, continued surveillance for amantadine/rimantadine-resistant influenza A viruses is warranted.

Real-time quantitative PCR analysis for alpha-thalassemia-1 of Southeast Asian type deletion in Taiwan.

Since homozygosity of the alpha-thalassemia-1 of Southeast Asian (SEA) type deletion results in hydrops fetalis, a novel protocol based on the real-time quantitating polymerase chain reaction (PCR) technique has been developed to quantify the intact and aberrant alpha-globin genes in adults. The ratio of the normal/SEA-bearing alpha-globin genes was expressed in cycle threshold (C(T)) values. Theoretically, a relative ratio of one to one was anticipated in individuals carrying the SEA type deletion. Twenty-five heterozygous and 20 normal cases were analyzed retrospectively with this protocol. Data showed that the CT values for the intact alpha-globin gene allele and the allele bearing the SEA type deletion in carriers were 28.74+/-1.49 and 26.46+/-2.05, respectively. Therefore, the ratio of normal/SEA type deletion-bearing alpha-globin genes in the carriers was 1.09+/-0.043. No ambiguous results were observed from other less common genotypes associated with alpha-thalassemia, such as the Philippine type deletion. Based on the results, we concluded that this protocol could provide a rapid method to mass screen carriers with alpha-thalassemia-1 of SEA type deletion in this region.

Characterization of serum and urinary chromogranin A by size exclusion chromatography: impact on calibrator selection and urinary assay.

Serum chromogranin A (CgA) is a useful marker for neuroendocrine tumors and is detectable in carcinomas at advanced stages. Elevated serum CgA is also an indicator of poor prognosis in prostate cancer and is useful for predicting the failure of hormonal therapy for prostate cancer patients. We found that CgA molecules with three different sizes could be detected in normal human serum. However, only the largest CgA molecule appears in patients with liver disease. Serum taken from cancer patients is composed predominantly of the middle-sized molecule, whereas the smallest CgA molecule was elevated in serum drawn from renal patients. Moreover, only the smallest CgA molecule was found in urine. We believe that the largest CgA molecule is metabolized by the liver, whereas the smallest CgA molecule is removed from the blood circulation via the kidney. Because the medium-sized CgA is the dominant molecule in both the cell medium of the tumor cell line SK-N-AS and sera from patients with malignant diseases, CgA from the cell medium was selected as the calibrator for the CgA ELISA assay. Our findings also suggest that it would not be possible to measure the urinary CgA to reflect the serum CgA concentration in order to detect pheochromocytoma among patients with hypertension.

Development of two ELISA for estrogen and progesterone receptor with sufficient sensitivity for fine needle aspirate and core biopsy.

The quantitative determination of estrogen and progesterone receptors (PR and ER) in breast tumor cytosol has been routinely performed in clinical laboratories to aid in the selection between hormonal and chemotherapy and also to predict prognosis. However, the small amount of tissue available from the increasingly popular fine-needle aspiration and core biopsies from breast cancer patients requires more sensitive immunoassays for receptor quantification. We have developed two sensitive immuno-assays for ER and PR on microplate with the use of recently available anti-ER and anti-PR antibodies of higher affinity and a powerful signal magnification agent, namely Amdex. The calibrator was a pooled breast tumor cytosol used as calibrator and calibrated against Abbott kits. The protein concentration of the cytosol and the upper normal cutoffs for our assays were reduced to approximately 0.2 mg/mL and 3 fmol/0.2 mg/mL, respectively. Both assays have sensitivities close to 1 fmol/mL, which are sufficiently sensitive for the receptor quantification in fine-needle aspiration biopsies and cord biopsies of breast tumor.

Genetic analysis of enterovirus 71 isolated from fatal and non-fatal cases of hand, foot and mouth disease during an epidemic in Taiwan, 1998.

A large scale outbreak of hand-foot-and-mouth disease (HFMD) occurred in Taiwan in 1998, in which more than 80 children died of shock syndrome with pulmonary edema/hemorrhage. Enterovirus 71 was implicated as the cause of this outbreak. In order to understand the virological basis responsible for mortality on this scale, nucleotide sequences of VP1 that is important for serotypic specificity, and the 5'-non-coding region (5'-NCR) that is important for replication efficiency, were analyzed comparatively. Phylogenetic analysis of both VP1 and 5'-NCR of nine EV71 isolates derived from specimens of fatal patients and seven isolates derived from uncomplicated HFMD patients showed that all but one isolate fell into genotype B. The one distinct isolate from a case of uncomplicated HFMD belonged to genotype C that was clustered along with one isolate from Taiwan in 1986. Complete sequence analysis of two selected isolates, one from the spinal cord of a fatal case and one from the vesicle fluid of a patient with mild HFMD, confirmed a high degree (97-100%) of identity in nucleotide sequence throughout the entire genome, except focal regions of 3C and 3'-NCR where the nucleotide homology was 90-91%. The identity of the deduced amino acid sequence in the 3C region that encodes viral proteinase dropped further to 86%, a result of missense mutations at the first nucleotide position of many codons.

Elevated serum chromogranin A is detectable in patients with carcinomas at advanced disease stages.

Chromogranin A (CgA), a marker of neuroendocrine cells and an indicator for neuroendocrine differentiation, is associated with a poor prognosis when detected in tumor tissue, based on immunohistochemical techniques. We sought to determine whether it is possible to detect elevated serum CgA in patients with commonly occurring carcinomas of non-neuroendocrine origin. CgA was measured in both random and serial serum specimens, using a serum CgA assay developed in our laboratory. Elevated levels of serum CgA were detected in patients with carcinoma of the prostate, breast, ovary, pancreas, and colon. Serum CgA levels in patients with all types of carcinoma appeared to parallel the changes of serum dominant tumor markers and were found in sera containing highly elevated tumor markers. Based on these preliminary findings, perhaps we should monitor CgA, in addition to the routinely used tumor markers, during the treatment of patients with carcinomas to determine if CgA is useful as a prognostic marker in carcinomas other than prostatic cancer.

Expression of capsid [correction of caspid] protein VP1 for use as antigen for the diagnosis of enterovirus 71 infection.

To produce enterovirus 71 antigen for diagnostic purposes, the gene encoding the entire capsid protein VP1 was amplified by reverse transcription-polymerase chain reaction (RT-PCR), cloned and expressed in Escherichia coli as a poly-histidine fusion protein. Western blotting experiments with sera from patients with enterovirus 71 infection indicated that immunoglobulin G (IgG) and IgM antibodies bound to a single polypeptide VP1. According to these results, IgM anti-VP1 appeared in sera of patients with a symptomatic enterovirus 71 acute infection, whereas IgG anti-VP1 was present in sera of past infection. This finding suggests that detecting IgG and IgM immune responses against linear epitopes of recombinant VP1 is an effective means of determining the different phases of enterovirus 71 infection. In addition, sera containing coxsackie virus 16 (CA16) antibodies did not cross-react with the recombinant VP1 of enterovirus 71, despite the homology between VP1 proteins of both viruses. Comparison with reference PCR and neutralization assays showed these antibody tests to be appropriate for the serodiagnosis of enterovirus 71 infection.

Non-radioactive Southern hybridization for early diagnosis of alpha-thalassemia with southeast Asian-type deletion in Taiwan.

Alpha-thalassemia has been estimated to account for over 60% of hydrops fetalis cases in Taiwan. The most common genotypic lesion found in alpha-thalassemia-1 cases in Taiwan is deletion of a large segment of the alpha-globin gene cluster, termed the Southeast Asian-type deletion (-SEA/; further referred to as SEA-type deletion). Seven chorionic villus samples (CVS) from pregnancies of couples both heterozygous for SEA-type deletion were studied. Non-radioactive Southern-blot hybridization using the dig-alkaline phosphatase detection system was developed to fulfill this purpose. The results were compared with corresponding polymerase chain reaction (PCR) data to elucidate the effectiveness of these two protocols in the diagnosis of the SEA-type deletion. The data showed that of the seven CVS, three demonstrated a distinctive band pattern, indicating their homozygous status of SEA-type deletion, whereas two showed heterozygous patterns, and the other two were free of the deletion. Homozygosity of the deletion was confirmed by Southern-blot hybridization performed on DNA samples extracted from the abortus tissue. However, two of the three cases with SEA-type deletion showed heterozygous PCR results. Maternal cell contamination could be responsible for the artifacts in the PCR results, but the influence due to the contamination is minimal in non-radioactive Southern-blot hybridization. We concluded that PCR is suitable for screening of carrier adults with SEA-type deletion, and non-radioactive Southern hybridization is ideal for early prenatal diagnosis of the SEA-type deletion.

Comparison of enterovirus 71 and coxsackie-virus A16 clinical illnesses during the Taiwan enterovirus epidemic, 1998.

OBJECTIVES: To compare enterovirus 71 (EV 71) with coxsackievirus A16 (Cox A16) clinical illness in patients at Chang Gung Children's Hospital during Taiwan's enterovirus epidemic of 1998. METHODS: With the use of the immunofluorescence assay and neutralization test, 177 cases of EV 71 and 64 cases of Cox A16 illness were confirmed from April to September, 1998. The clinical signs and symptoms, complications and case fatality rates were compared. RESULTS: Three-fourths of the cases were younger than 3 years of age, and the ratio of males to females was 1.3 in the EV 71 group and 1.2 in the Cox A16 group. In the EV 71 group 120 (68%) cases were uncomplicated, including 94 cases of hand, foot and mouth disease and 15 cases of herpangina, and 57 (32%) cases had complications, including 13 (7.3%) cases of aseptic meningitis, 18 (10%) cases of encephalitis, 4 (2.3%) cases of polio-like syndrome, 8 (4.5%) cases of encephalomyelitis and 12 (6.8%) cases of fatal pulmonary edema. Fourteen (7.9%) patients died, including 12 cases of pulmonary edema and 2 cases of encephalitis; seven (4%) patients had sequelae. By contrast, 60 (94%) of the 64 cases of Cox A16 infection were uncomplicated and only 4 (6.3%) cases were complicated by aseptic meningitis; no fatalities or sequelae were observed. By multivariate analysis vomiting (P = 0.01) and fever higher than 39 degrees C plus lasting longer than 3 days (P = 0.02) were significantly more frequent in the EV 71 group. CONCLUSION: EV 71 illness is more severe with significantly greater frequency of serious complications and fatality than is illness caused by Cox A16.

Diagnosis of respiratory tract viruses in 24 h by immunofluorescent staining of shell vial cultures containing Madin-Darby Canine Kidney (MDCK) cells.

Nine hundred and seventy-eight clinical specimens were examined taken from patients with respiratory tract viruses (RV)-like syndrome between November 1996 and July 1998. The study was undertaken to evaluate the effectiveness of centrifuge-enhanced shell vial cultures (SVC) containing Madin-Darby Canine Kidney (MDCK) cells, combined with immunofluorescent (IF) staining in 24 h. This technique rapidly detects and identifies respiratory tract viruses. The conventional tube culture system with multiple cell lines would ordinarily detect RV within 3-30 days. The SVC/IF method using single cell line (MDCK cells) allowed detection of 81.5% of influenza A virus, 72% of parainfluenza virus, 82.6% of respiratory syncytial virus (RSV) and 79.6% of adenovirus in 24 h.

High prevalence of TT virus infection in healthy children and adults and in patients with liver disease in Taiwan.

A newly identified DNA virus, named TT virus (TTV), was found to be related to transfusion-associated hepatitis. We conducted the following experiments to evaluate its pathogenic role in liver disease and potential modes of transmission. We used PCR to detect TTV DNA in serum. The rates of TTV viremia in 13 patients with idiopathic acute hepatitis, 14 patients with idiopathic fulminant hepatitis, 22 patients with chronic hepatitis, and 19 patients with cirrhosis of the liver were 46, 64, 55, and 63%, respectively, and were not significantly different from those in 50 healthy control subjects (53%). PCR products derived from seven patients with liver disease and three healthy controls were cloned and then subjected to phylogenetic analyses, which failed to link a virulent strain of TTV to severe liver disease. TTV infection was further assessed in an additional 148 subjects with normal liver biochemical tests, including 30 newborns (sera collected from the umbilical cord), 23 infants, 16 preschool children, 21 individuals of an age prior to that of sexual experience (aged 6 to 15 years), 15 young adults (aged under 30 years), and 43 individuals older than 30 years. The rates of TTV viremia were 0, 17, 25, 33, 47, and 54%, respectively. These findings suggest that TTV is transmitted mainly via nonparenteral daily contact and frequently occurs very early in life and that TTV infection does not have a significant effect on liver disease.

Lymphangiosis as a predictor of outcome in patients with primary diffusely infiltrative adenocarcinoma of the colon and rectum.

OBJECTIVE: To investigate the relationships between outcome and clinicopathological factors, DNA flow cytometrical characteristics, and postoperative adjuvant therapy in patients with primary diffusely infiltrative colorectal adenocarcinoma. DESIGN: Inception cohort study. SETTING: A medical center that offers a mixture of primary, secondary, and tertiary care services. PATIENTS: Among 7035 patients undergoing resection of primary colorectal adenocarcinoma from 1980 to 1996, 37 patients with a pathological diagnosis of primary diffusely infiltrative tumor were selected. All patients had received regular follow-up until February 28, 1998, or until death. MAIN OUTCOME MEASURES: Cancer-specific survival compared by log-rank test and Cox regression model. RESULTS: Univariate analyses revealed tumor stage (stages II-III vs. stage IV, P = .01) and severity of lymphangiosis (absent/mild vs. moderate/severe, P = .04) were significant in predicting outcome. A proliferative index of greater than 20% was insignificant (P = .08) in predicting outcome. In a Cox regression model, TNM stage and lymphangiosis were independently correlated with a worse outcome. When compared with tumors having less severe lymphangiosis, the odds ratio of death due to cancer in cases of tumors with moderate to severe lymphangiosis was 2.4 (95% confidence interval, 1.0-5.6; P = .05). CONCLUSION: Lymphangiosis and TNM stage were independently predictive of outcome in patients with primary diffusely infiltrative colorectal cancer.

Increased absolute number but not proportion of gamma/delta T-lymphocytes in the bronchoalveolar lavage fluid of patients with active pulmonary tuberculosis.

SETTING: The proportions and absolute cell count of gamma/delta T-lymphocytes in the peripheral blood of patients with pulmonary tuberculosis (PTB) remains controversial. Since PTB is an infections airway disease, bronchoalveolar T-lymphocytes should be a better indicator of local immune T-cell reaction after TB infection than peripheral blood T-lymphocytes. OBJECTIVE: To quantitate the absolute cell count and proportions of gamma/delta T-lymphocytes in the bronchoalveolar lavage fluid (BALF) of patients with active PTB. DESIGN: Bronchoalveolar lavage (BAL) and analysis of lymphocytes in the BALF was performed in 25 patients with active PTB and 16 normal controls. All of the patients were negative for HIV infection and none was immunocompromised. BALF and blood were prepared for cell differential count and flow cytometry analysis using monoclonal antibodies CD3, CD4, CD8, CD25, HLA-DR and gamma/delta as well as alpha/beta T-lymphocyte receptors. RESULTS: The number of cells per volume of recovered BALF was significantly higher in the patients with active PTB than in normal controls. BALF from active PTB patients also showed increased percentage of lymphocytes and neutrophils. The absolute number of total lymphocytes, CD3+ lymphocytes and CD3+ gamma/delta T-lymphocytes were significantly higher in the BALF, but not in the blood, of patients with TB, however, the proportions of CD3+ gamma/delta T-lymphocytes in BALF of patients with TB was comparable to that of normal controls. gamma/delta T-lymphocytes in the BALF rarely expressed CD4, CD25, and HLA-DR in both groups. CONCLUSION: These results suggest that gamma/delta T-lymphocytes are not the major subpopulation of CD3+ lymphocytes in the BALF that react to mycobacterial infection in the patients with clinically established active TB.