Injury induces deficient interleukin-12 production, but interleukin-12 therapy after injury restores resistance to infection

OBJECTIVE: To assess at serial intervals the production of interleukin-12 (IL-12) by monocytes/macrophages from the peripheral blood of injured patients and control subjects, and using a mouse model to confirm human findings and explore the effectiveness of low-dose IL-12 therapy in restoring resistance to infection after injury. SUMMARY BACKGROUND DATA: Serious injury is associated with loss of function of the T helper 1 lymphocyte phenotype, but little is known about IL-12 production in injured patients. The authors previously reported that early, moderate-dose IL-12 therapy in a mouse model of burn injury restored resistance to a later infectious challenge (cecal ligation and puncture, CLP). However, the efficacy of clinically relevant low-dose IL-12 therapy carried out to or beyond the time of septic challenge remains to be tested. METHODS: Peripheral blood mononuclear cells (PBMCs) and adherent cells were obtained from 27 patients with major burns or traumatic injury and 18 healthy persons and were studied at serial intervals for IL-12 production stimulated by bacterial lipopolysacharide (LPS). PBMCs from 18 of the same patients were studied for IL-10 production as well. IL-12 production by adherent cells from the spleens of burn or sham burn mice was studied at serial intervals after injury to confirm the human findings. Low-dose IL-12 or vehicle was given every other day to groups of burn and sham burn mice, which were then challenged with CLP on day 10, and survival was determined. Finally, spleens were harvested from burn or sham burn animals receiving low-dose IL-12 or vehicle after CLP. After splenic cellularity was determined by hemocytometer, splenocytes were cultured and production of tumor necrosis factor-alpha, interferon-gamma, and IL-10 were assessed by immunoassay. RESULTS: Adherent cells from patients' PBMCs produced significantly less IL-12 than normal PBMCs after injury, reaching a nadir 8 to 14 days after injury. Stimulation of whole PBMCs by LPS indicated that at 8 to 14 days after injury, IL-12 production by PBMCs was significantly lower and IL-10 production was significantly higher than that of PBMCs from healthy persons. Low-dose IL-12 therapy significantly increased survival after CLP. Splenocytes from burn mice treated with IL-12 had significantly increased production of TNF-alpha and IF-beta, both before and after CLP, when compared with vehicle-treated burn animals. IL-10 production by bum splenocytes remained high after IL-12 treatment. Splenic cellularity increased after IL-12 treatment in burn mice. CONCLUSION: The capacity to produce IL-12 by adherent cells of the monocyte/macrophage lineage is significantly reduced after serious injury in humans and in a mouse burn model. In humans, there is a reciprocal relation between diminished IL-12 production and increased IL-10 production at approximately 1 week after injury. Low-dose IL-12 therapy in the mouse burn model markedly increased survival after a septic challenge, even when treatment was carried beyond the onset of sepsis. Low-dose IL-12 treatment in the mouse increased production of proinflammatory mediators important in host defense and at the same time maintained or increased production of IL-10, an important antiinflammatory cytokine.     

Major injury induces increased production of interleukin-10 by cells of the immune system with a negative impact on resistance to infection.

OBJECTIVE: The purpose of this study was to compare the production of interleukin-10 (IL-10) by peripheral blood mononuclear cells (PBMC) from injured patients and control subjects to determine the responsible cell types and to relate IL-10 production to the occurrence of sepsis. A mouse model of burn injury was used to confirm the human findings and to assess the importance of IL-10 in the lowered resistance to infection after injury. SUMMARY BACKGROUND DATA: Severe injury is associated with depressed immune responses. Although IL-10 is known to inhibit several aspects of immune reactivity, the role of IL-10 in postinjury immune suppression remains controversial. METHODS: Peripheral blood mononuclear cells from 14 burn and 12 trauma patients and 16 healthy individuals were studied at serial intervals for IL-10 production stimulated by a T-cell mitogen, phytohemagglutinin, and by bacterial lipopolysaccharide. To determine the source of IL-10, CD4+ and CD8+ lymphocyte subsets were obtained by selective depletion of PBMC with antibody-coated magnetic beads and were stimulated by anti-CD3 antibody to induce IL-10 secretion. In addition, IL-10 production by patients' PBMC in the first 10 days after injury was assessed for correlation with subsequent septic events. Anti-CD3-stimulated IL-10 production also was determined for CD4- and CD8-enriched lymphocyte subsets obtained by antibody and complement depletion of splenocytes harvested from groups of burn and sham burn mice at day 10 after injury, the time of maximal susceptibility to a septic challenge, cecal ligation and puncture (CLP). Finally, to test the importance of IL-10 in immune suppression in vivo, groups of burn and sham burn mice were treated with anti-IL-10 monoclonal antibody or control immunoglobulin G (IgG) on days 1 and 3 postinjury and were observed for survival after CLP on day 10. RESULTS: Patients' PBMC produced significantly more IL-10 than did controls' PBMC 7 to 14 days after injury. Patients' CD4+ (T-helper) but not CD8+ (T-cytotoxic) lymphocytes also showed increased IL-10 production versus those of control subjects early after injury. Increased PBMC IL-10 production in the first 10 days postinjury correlated significantly (p < 0.05) with subsequent septic events. Burn mouse CD4-enriched but not CD8-enriched splenocytes produced more IL-10 than did sham burn splenocyte subsets on day 10 after injury. Burn mice treated with anti-IL-10 antibody but not with control IgG had significantly increased survival after CLP. CONCLUSION: Serious injury in humans and in a mouse burn model is followed by increased stimulated production of IL-10 by cells of the immune system. The CD4+ T-helper cells appear to be a major source of IL-10 after injury. In injured patients, increased IL-10 production is correlated with subsequent septic events, and in the burn mouse, IL-10 appears to induce decreased resistance to infection.     

A mechanism of interleukin-12 unresponsiveness associated with thermal injury

An unresponsive state for the production of interleukin-12 (IL-12) is commonly observed in animals and patients with severe thermal injuries. In the present study, the participation of corticosteroids, prostaglandin E(2) (PGE(2)), and type 2 cytokines, which appeared in association with thermal injury, on the burn-associated IL-12 unresponsiveness was studied. These substances have been described as inhibitors of IL-12 production. Less than 20 pg/ml serum IL-12 was produced in thermally injured mice (TI-mice) after stimulation with lipopolysaccharide (LPS), while 1037 pg/ml IL-12 was detected in sera of unburned mice equally stimulated with LPS. Almost complete restoration of the impaired IL-12 production was witnessed in TI-mice after treatment with soluble IL-4 receptor (50 ng/mouse, 2 h and 2 days after thermal injury). However, IL-12 was not induced by LPS stimulation in TI-mice treated with an inhibitor of PGE(2) (indomethacin, 0.1-5 mg/kg) or an inhibitor of corticosteroid production (ketoconazole, 10 mg/kg). LPS-stimulated IL-12 production was also impaired in normal mice inoculated with burn-associated type 2 T cells. In addition, in the presence of 1 microg/ml LPS, naive macrophages cocultured with burn-associated type 2 T cells did not produce IL-12 in their culture fluids, while IL-12 was produced by LPS-stimulated naive macrophages that were cocultured with naive splenic CD8(+) T cells. These results suggest that the IL-12-unresponsive state demonstrated in TI-mice is associated mainly with type 2 cytokines released from burn-associated type 2 T cells.     

Frequency, specificity, and phenotype of clonally growing human alloreactive interleukin-2-secreting helper T lymphocyte precursors

Limiting dilution cultures were performed to detect allospecific IL-2-secreting helper T lymphocyte precursors (HTL-p) among human peripheral blood mononuclear cells, E-rosette-purified (E+) and cell-sorter-separated CD4+/8- as well as CD4-/8+ T cell subsets. Split-well cultures were set up prior to restimulation to assess the antigen specificity of the response. Frequencies of alloreactive IL-2-secreting HTL-p in fully HLA-mismatched responder/stimulator cell combinations ranged from 1/200 to 1/900 (among PBMNC), from 1/50 to 1/301 (among E+ T cells), from 1/36 to 1/220 (among CD4+ T cells), and from 1/38 to 1/450 (among CD8+ T cells). Allospecificity of effector T cells was demonstrated by a strong decline of frequencies obtained after restimulation against unrelated third-party antigens. In clonal segregation analysis, the vast majority of IL-2-secreting progeny (80-90%) were exclusively specific for the original stimulating alloantigen. Finally, the allele specificity of human alloreactive HTL-p was revealed by comparing frequency estimates obtained after restimulation with partially identical stimulator/third-party antigen combinations.     

Prolongation of allograft survival by Nippostrongylus brasiliensis is associated with decreased allospecific cytotoxic T lymphocyte activity and development of T cytotoxic cell type 2 cells

BACKGROUND: We have demonstrated that infection with Nippostrongylus brasiliensis (Nb), which induces strong type 2 responses, prolongs kidney allograft survival in rats. Here, we confirm that this effect is not species-specific and address immune modulation in allospecific T-cell responses mediated by nematode infection. METHODS: C57BL/6 mice were injected with Nb or phosphate-buffered saline. Four days later, mice were transplanted with BALB/c hearts and graft survival was assessed. In other experiments, Nb-infected mice were immunized with BALB/c spleen cells and allospecific T-cell responses were determined in vitro. RESULTS: In this study, we show that Nb prolongs cardiac allograft survival in mice. Further, spleen T cells from Nb-infected, allo-immunized mice exhibit reduced allospecific cytotoxic T-lymphocyte activity. In contrast, allospecific proliferation of T cells in the mixed lymphocyte reaction was not reduced by Nb, ruling out immunosuppression as the mechanism of Nb-induced allograft survival. Nb infection induced IL-4 and IL-6 and inhibited IFN-gamma production by T cells in response to allo-antigen. Furthermore, anti-IL-4 treatment reduced allospecific T-cell proliferation from Nb-infected but not control mice, indicating that type 2 allospecific T cells develop in the presence of Nb. We also double-stained T cells for CD8 and IL-4 and showed that Nb induces an 8-fold increase in Tc2 cell numbers. CONCLUSIONS: These results are consistent with a hypothesis that Nb mediates prolongation of allograft survival through induction of type 2 immunity, including the development of regulatory Tc2 cells, and subsequent inhibition of allospecific cytotoxic T-lymphocyte activity.     

Effect of histamine on intercellular adhesion molecule-1 expression and production of interferon-gamma and interleukin-12 in mixed lymphocyte reaction stimulated with interleukin-18

BACKGROUND: Interleukin (IL)-18 was identified as an interferon (IFN)-gamma-inducing factor and was demonstrated to up-regulate the expression of intercellular adhesion molecule (ICAM)-1 on human monocytes. In organ transplantation, elevation of plasma IL-18 levels has been reported during acute rejection. In the present study, we examined the effect of IL-18 on human mixed lymphocyte reaction (MLR), an in vitro model of acute rejection after organ transplantation. We also investigated the modulatory effects of histamine on IL-18 action because histamine has been demonstrated to be a modulator of IL-18 effect and a mediator of inflammation. METHODS: We measured the expression of ICAM-1 on human monocytes in MLR in the presence or absence of IL-18 by flow cytometer and determined the associated production of IFN-gamma and IL-12 by ELISA. The modulatory effects of histamine and the relevant histamine receptor subtypes were characterized pharmacologically. RESULTS: The expression of ICAM-1 on monocytes in MLR was markedly enhanced by the addition of IL-18 in a concentration- and time-dependent manner. In parallel to ICAM-1 up-regulation, IL-18 significantly enhanced the production of IFN-gamma and IL-12 in MLR. Histamine concentration-dependently inhibited ICAM-1 expression and cytokine production in MLR stimulated with IL-18, whereas histamine alone did not show any effects on these responses in the absence of IL-18. The effects of histamine on both ICAM-1 expression and cytokine production were mimicked by the selective H2-receptor agonists 4-methylhistamine and dimaprit and were antagonized by the H2-receptor antagonist famotidine but not by H1- and H3-receptor antagonists. CONCLUSION: IL-18 strongly enhanced human MLR with respect to ICAM-1 expression and cytokine production. The fact that histamine could inhibit the IL-18-stimulated MLR implies that immunomodulation by histamine and selective H2-receptor agonists may have an important role in future immunosuppressive strategies.     

Local production of interleukin-8 is associated with nosocomial pneumonia.

One hundred five (70%) of 151 patients hospitalized in the intensive care unit and undergoing mechanical ventilation had bronchial secretions that tested positive for interleukin-8 within 36 hours of admission. Arterial blood, mixed venous blood, and urine collected simultaneously all tested negative, except for 11 patients admitted with intra-abdominal septic foci. The presence of interleukin-8 in the pulmonary air space early in the course of hospitalization was significantly associated with patients with multiple injuries, the need for greater ventilatory support, the occurrence of pulmonary dysfunction, and a 66% incidence of nosocomial bacterial pneumonia. We conclude that the early local production of interleukin-8 in the lungs is an early marker of pulmonary injury and may be involved in the pathogenesis of nosocomial bacterial pneumonia.     

Acute gastric mucosal injury associated with the systemic administration of interleukin-4.

METHODS. Seventy-three patients with advanced malignancy were treated with the recombinant lymphokine interleukin-4 either as the sole immunotherapeutic reagent or in combination with recombinant interleukin-2. RESULTS. Twelve of 84 courses of therapy were complicated by gastroduodenal erosion or ulceration. Three of these courses were associated with significant bleeding, which required multiple red blood cell transfusions and endoscopic therapy. No treatment-related deaths occurred. Eleven of 57 courses administered with concomitant indomethacin and 11 of 62 courses administered with ranitidine resulted in gastroduodenal mucosal injury. No acute change in gastric acid output occurred after one dose of interleukin-4 in patients prospectively evaluated with an indwelling nasogastric tube. An intravenous ranitidine infusion appropriately reduced acid output in these patients. In contrast, we have treated over 650 patients with interleukin-2 and indomethacin without interleukin-4, none of whom developed signs or symptoms of gastroduodenal ulceration. CONCLUSIONS. These observations suggest that systemically administered cytokines may exert an effect on the integrity of the gastroduodenal mucosa.     

Changes in production of interleukin-1 and interleukin-2 associated with obstructive jaundice and biliary drainage in patients with gastrointestinal cancer

Increased susceptibility to infection in patients with obstructive jaundice has been well documented in vitro and in vivo. Nevertheless, an underlying mechanism for immunocompromise in these patients has not been identified. This study was undertaken to evaluate the production of two important immunoregulatory molecules, interleukin-1 (IL-1) and interleukin-2 (IL-2), by peripheral blood mononuclear cells in cancer patients with obstructive jaundice before and after percutaneous transhepatic biliary drainage (PTBD). After decompression with PTBD, IL-1 and IL-2 production was significantly increased (IL-1: from 7.9 +/- 4.9 to 13.9 +/- 4.9 U/ml, p less than 0.05; IL-2: from 8.8 +/- 4.9 to 14.1 +/- 6.5 U/ml, p less than 0.05). There was a positive correlation between IL-1 and IL-2 production (r = 0.424, p less than 0.05). The production of both interleukins correlated negatively with serum total bilirubin level (IL-1 r = -0.478, p less than 0.05; IL-2: r = -0.482, p less than 0.05) and positively with high-density lipoprotein cholesterol in serum (IL-1: r = 0.505, p less than 0.01; IL-2: r = 0.494, p less than 0.05). IL-2 production also correlated positively with serum albumin levels (r = 0.511, p less than 0.01). These results suggest that hyperbilirubinemia and abnormal lipid metabolism may be associated with impaired interleukin production, which may result in an increased susceptibility to infection during obstructive jaundice.     

Vibrio infection associated with finning injury of the hand

BACKGROUND: Penetrating injury by fish fins is common and often overlooked. Vibrio spp. are known worldwide for their virulence, quickly causing soft-tissue infection and lethal septicaemia. Vibrio infection following finning injury is rare, but can result in devastating complications in susceptible individuals. AIM: To elucidate the clinical significance of such injury. METHOD: Between July 2003 and September 2005, nine cases of Vibrio infection caused by finning injury to the hand were retrospectively reviewed. Clinical data, including skin presentations, treatment course and outcomes, were collected and reviewed. RESULTS: In our group of nine patients, seven had concurrent hepatoma, diabetes mellitus, cirrhosis, chronic renal insufficiency or the effects of long-term steroid use; three had wound infections manifested by cellulitis or tenosynovitis and six had life-threatening necrotising soft-tissue infections. Vibrio spp. were identified from the wound (n=4), blood (n=1), and both (n=4). Symptoms appeared within a few hours to 3 days after injury, with 50% of patients developing symptoms within 24h; three patients were hypotensive upon admission; one patient received antibiotic therapy only and eight required emergency fasciotomy. All patients survived and none required amputation. The mean hospital stay was 23.2 days. CONCLUSIONS: Vibrio infections after finning injury can produce bacteraemic necrotising soft tissue-infections, especially in individuals with a systemic illness. Health education should include a recommendation to wear protective gloves while handling fish. Early antibiotic and surgical treatment can avoid potentially life-threatening complications.     

白细胞介素-2基因转染联合特异性细胞毒性T淋巴细胞对裸鼠肝癌的影响

目的 观察白细胞介素-2(IL-2)基因转染联合特异性细胞毒性T淋巴细胞(sCTL)治疗裸鼠肝癌的疗效.方法 荷肝癌裸鼠6只,随机分为pORF-hIL-2质粒转染实验组和对照组,检测血浆白细胞介素(IL)-2的浓度.荷肝癌裸鼠24只,随机分成对照组、IL-2基因治疗组、sCTL治疗组和IL-2基因联合sCTL治疗组,观察肿瘤体积变化及生存时间.结果 实验组血清IL-2浓度在转染后第3天到达顶峰135.3 ng/L,持续15 d,对照组检测不到IL-2;各组肿瘤体积(治疗15 d后)变化为(262.27±78.30)、(151.65±63.93)、(27.16±27.69)、(-63.32±16.92)mm3;中位生存时间20、27.5、60、90 d;差异均有统计学意义(P＜0.01).结论 联合应用IL-2基因转染和sCTL细胞治疗裸鼠的HepG2移植瘤,可以明显地提高抗瘤效应,表现协同增效作用.     

IL-2基因转染联合CTL杀伤肝癌细胞的体外试验研究

目的 通过研究白细胞介素-2（IL-2）基因转染联合特异性细胞毒T淋巴细胞（CTL）对人肝癌细胞（HepG2）的体外杀伤试验，探讨一种肝癌的免疫基因治疗方法。方法 采用多聚阳离子脂质体作载体，将IL-2基因转染至HepG2细胞。通过检测IL-2浓度变化，观察转染效率。用MTT法检测其联合特异性CTL细胞对HepG2细胞的杀伤作用。结果 转染IL-2基因至HepG2细胞的体外最佳转染比例为5：1，其可持续分泌IL-230d。分泌高峰在转染后第2—3天。联合特异性CTL细胞，对HepG2细胞显示明显的协同杀伤效应。结论 经转染IL-2基因后，HepG2细胞分泌IL-2可达30d，可以有效地维持特异性CTL细胞的活力，两者联合，对杀伤肝癌细胞有明显的协同效应。     

Flow cytometric measurement of rat lymphocyte subpopulations after burn injury and burn injury with infection

Increased infection rates in burned patients may result from a disproportionate increase in the suppressor subpopulations. Measurement of lymphocyte subpopulations is difficult in burned patients because gradient-purified cells are contaminated by nonlymphoid cells. The accuracy of flow cytometric subpopulation analysis was improved by restricting (gating) the analysis to cells with light-scatter intensity typical of lymphocytes. Blood was obtained 48 hours after burn from rats receiving no burns, 30% scald burns, or burns seeded with Pseudomonas aeruginosa to induce infection. Subpopulations were identified by monoclonal antibodies to T-lymphocyte antigens. Gating increased the values obtained for most subpopulations, but the relative differences between groups were unchanged. Burned and infected animals, but not animals burned only, had a decreased ratio of helper to suppressor lymphocytes (HSR) relative to control. A decreased HSR correlated with sepsis, but not with infection susceptibility. This suggests that a decrease in HSR may be a result of infection rather than a cause of susceptibility to infection.     

Pre-transplant elevations of interleukin-12 and interleukin-10 are associated with acute rejection after renal transplantation

BACKGROUND: Methods for predicting patients at higher risk for rejection before transplantation may help improve outcomes. We hypothesized that pre-transplant elevations of serum interleukin-12 (IL-12), a pro-inflammatory cytokine, would predict acute rejection, while pre-transplant IL-10, an immunoregulatory cytokine, would be down-regulated in patients subsequently experiencing acute rejection. MATERIALS AND METHODS: Thirty patients experiencing acute rejection after cadaveric renal allograft transplantation and a control group of 30 patients, undergoing the same procedure but without the occurrence of rejection, were identified. Serum samples taken before transplantation from each patient were then analyzed quantitatively for IL-12 and IL-10 using ELISA assays. RESULTS: The mean pre-transplant serum IL-12 level was higher in patients who subsequently underwent acute rejection vs. those who did not (181 +/- 143 pg/mL vs. 81.2 +/- 71.5 pg/mL, respectively, p = 0.007). Unexpectedly, pre-transplant serum IL-10 levels were also elevated in patients who underwent rejection (559 +/- 293 pg/mL vs. 332 +/- 163 pg/mL, respectively, p = 0.002). Multivariate analysis demonstrated that elevations of IL-12 and IL-10 were independent risk factors for rejection when adjusted for confounding variables. CONCLUSIONS: Pre-transplant elevations of IL-12 and, unexpectedly, IL-10 are associated with acute rejection after cadaveric renal transplantation and may be useful in predicting which patients are at increased immunological risk at the time of transplantation. Further studies are necessary to assess the role of occult systemic inflammation in contributing to poor outcomes after transplantation.     

Increased norepinephrine production associated with burn injuries results in CCL2 production and type 2 T cell generation

The susceptibility of thermally injured mice (TI-mice) to various infections is markedly influenced by burn-associated type 2 T cell responses, which are common with severe thermal injuries. Previously, we have reported that CC chemokine ligand 2/monocyte chemoattractant protein-I (CCL2) is produced in mice within 1 day of thermal injury, and the subsequent development of burn-associated type 2 T cell responses are triggered by this chemokine produced early after thermal injury. In this study, influence of norepinephrine (NE) on CCL2 production in mice early after thermal injury (TI) was investigated. Peripheral blood mononuclear cells (PBMC) and peritoneal macrophages (PMphi) from TI-mice produced CCL2, but the same cell preparations from normal mice did not. This chemokine was not produced by PBMC and PMphi from TI-mice previously treated with 6-hydroxydopamine (6-OHDA), which destroys sympathetic nerve termini. NE production was increased in circulation of TI-mice, and treatment of TI-mice with 6-OHDA resulted in the inhibition of NE secretion. When PBMC from normal mice were treated with NE, they acquired the ability to produce CCL2. Splenic T cells from TI-mice produced IL-4 into their culture fluids, while the cytokine was not produced by splenic T cells from TI-mice previously treated with 6-OHDA. These results indicate that NE may have an important role on early CCL2 production and the subsequent development of burn-associated type 2 T cell responses.     

Changes in T lymphocyte subsets following injury. Assessment by flow cytometry and relationship to sepsis.

The increased susceptibility of severely injured patients to infection and death from sepsis has been attributed to abnormalities in cell-mediated immunity. The authors therefore assessed the relative number of peripheral blood T helper cells and T suppressor/cytotoxic cells and total T lymphocytes identified by the monoclonal antibodies (McA) OKT4, OKT8, and OKT3, respectively, in 25 patients with burns from 5 to 85% total body surface area (TBSA) (mean: 40%) and 21 patients with nonthermal injuries (mean Injury Severity Score (ISS): 21.4). Patients were compared to 21 healthy controls. Cells reacting with the McA were detected by flow cytometry, which enabled the examination of a population of cells the size of T lymphocytes, excluding larger contaminating cells that might bind the McA. Patients with burns of 30% TBSA or greater had a significant reduction (p less than or equal to 0.05) in OKT3+ cells up to 50 days post-burn. Both septic and nonseptic burn patients had reduced numbers of OKT3+ cells, as did patients after nonthermal injury, suggesting that this reduction was due to the injury itself. Patients with smaller burns (less than 30% TBSA) as a group did not have reduced OKT4+ cells, whereas those with larger burns showed significant reductions in OKT4+ cells (P less than or equal to 0.05) at 0 to 5, 6 to 10, 11 to 20, 21 to 30, and 41 to 50 days post-burn. Seven burn patients who became septic 10 days post-burn or later had significantly lower OKT4+ cells within 10 days of injury (mean: 33.75% +/- 7.4 SEM) than 10 patients who remained free of sepsis (mean: 42.2% +/- 5.4, p = 0.004). Patients with uncomplicated nonthermal injuries failed to show any significant reduction in OKT4+ cells. Following thermal injury, a reduction in OKT8+ cells was observed up to 10 days in patients with burns less than 30% TBSA, and up to 20 days in patients with larger burns. In both groups, at no time were increased OKT8+ cells found to correlate with clinical events. In patients with nonthermal injury, OKT8+ cells generally remained near the normal range.     

Peripheral T-cell lymphoma with Reed-Sternberg-like cells of B-cell phenotype and genotype associated with Epstein-Barr virus infection.

We report three cases of nodal peripheral T-cell lymphoma (PTCL) with Reed-Sternberg-like (RS-like) cells of B-cell pheno- and/or genotype. Histologic analysis in all cases revealed diffuse nodal effacement by atypical lymphoid cells of variable size. Two of the three cases had features of angioimmunoblastic T-cell lymphoma (AILT). Large mononuclear and binucleated cells with prominent eosinophilic nucleoli and abundant cytoplasm resembling classic RS cells and mononuclear variants were scattered throughout all biopsies. The lymphoma cells in the three cases were of T-cell lineage (CD3+, CD43+, and CD45RO+). The RS-like cells from all cases were CD30 and CD15 positive. In contrast to the neoplastic T cells, the RS-like cells lacked all T-cell markers and in two cases were positive for CD20. Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) and EBER 1 (2/2) were detected in the RS-like cells in all cases. The neoplastic T cells were negative for EBV. Polymerase chain reaction (PCR) analysis demonstrated clonal rearrangements of the T-cell receptor gamma chain gene in the three cases. PCR analysis of microdissected RS-like cells for immunoglobulin heavy chain gene rearrangements in cases 1 and 3 showed an oligoclonal pattern. The presence of RS-like cells in PTCL represents a diagnostic pitfall, because in one case this observation led to a misdiagnosis of Hodgkin's disease (HD). The oligoclonal expansion of EBV-infected cells may be related to underlying immunodeficiency associated with T-cell lymphomas and AILT in particular. This phenomenon may provide the basis for some cases of Hodgkin's disease after T-cell lymphomas and suggests that they are clonally unrelated neoplasms. The expression of LMP1 appears to be crucial for the immunophenotype and probably for the morphology of the RS and RS-like cells appearing in diverse lymphoid malignancies, including HD, chronic lymphocytic leukemia, and PTCL.