Angela Capobianchi

Cytomegalovirus infection in inflammatory bowel disease patients undergoing anti-TNFα therapy

BACKGROUND Cytomegalovirus infection and disease is associated with poor prognosis and steroid refractoriness in inflammatory bowel disease patients. The unfavourable effect of steroids and immunosuppressive therapy on CMV infection is well known but few data are available concerning anti-TNFalpha therapy (Infliximab). Aim of the study was to evaluate the presence and severity of CMV infection and disease in Infliximab-treated IBD patients. PATIENTS AND METHODS The severity of active CMV infection and disease was assessed in 11 consecutive patients with ileocolonic/colonic disease and 4 patients with ulcerative colitis before and after a standard 3-infusion course of Infliximab. Active CMV infection was evaluated by serology and diagnosed by means of pp65-antigenemia (pp65 AG), and quantification of CMV DNA isolated from biopsy specimens of colonic tissue. CMV disease was assessed on haematoxylin/eosin-stained colonic biopsies and immunohistochemical stains. RESULTS Of the 11 patients, nine were CMV seropositive. As far as concerns CMV infection, only one patient had positive pp65 AG, before and after Infliximab. CMV DNA was detected in the colonic biopsies of three patients. In 2, CMV DNA persisted also after therapy with 410 and 1300 copies/microg of DNA, respectively, albeit with no evidence of worsening of the colonic disease. In the remaining patient, CMV DNA load became undetectable. Conventional histology and immunohistochemical stains were negative for CMV in all the patients, without evidence of CMV disease. CONCLUSIONS Active CMV infection did not progress to disease following Infliximab therapy. Although these preliminary observations require confirmation, the response to Infliximab therapy does not appear to be influenced by, or influence the course of, CMV infection/disease.

STAT3 activation by KSHV correlates with IL-10, IL-6 and IL-23 release and an autophagic block in dendritic cells

Kaposiss sarcoma associated herpesvirus (KSHV) has been reported to infect, among others, monocytes and dendritic cells DCs impairing their function. However, the underlying mechanisms remain not completely elucidated yet. Here we show that DC exposure to active or UV-inactivated KSHV resulted in STAT3 phosphorylation. This effect, partially dependent on KSHV-engagement of DC-SIGN, induced a high release of IL-10, IL-6 and IL-23, cytokines that in turn might maintain STAT3 in a phosphorylated state. STAT3 activation also correlated with a block of autophagy in DCs, as indicated by LC3II reduction and p62 accumulation. The IL-10, IL-6 and IL-23 release and the autophagic block could be overcome by inhibiting STAT3 activation, highlighting the role of STAT3 in mediating such effects. In conclusion, here we show that STAT3 activation can be one of the molecular mechanisms leading to KSHV-mediated DC dysfunction, that might allow viral persistence and the onset of KSHV-associated malignancies.

A prospective study comparing quantitative Cytomegalovirus (CMV) polymerase chain reaction in plasma and pp65 antigenemia assay in monitoring patients after allogeneic stem cell transplantation

BackgroundLow levels of Cytomegalovirus (CMV) viral load are frequently detected following allogeneic stem cell transplantation (SCT) and CMV disease may still develop in some allogeneic SCT patients who have negative pp65-antigenemia (pp65-Ag) or undetectable DNA. Pp65Ag is a sensitive method to diagnose CMV infection. Quantitative CMV-DNA PCR assay in plasma has been proposed to monitor CMV infection in SCT patients. We evaluated the clinical utility of pp65Ag and PCR assay in plasma of SCT recipients.MethodsIn a prospective longitudinal study, 38 consecutive patients at risk of CMV infection (donor and/or recipient CMV seropositive) were weekly monitored for CMV infection by both quantitative CMV-PCR in plasma (COBAS AMPLICOR CMV MONITOR) and pp65 Ag, during the first 100 days after SCT.ResultsA total of 534 blood samples were simultaneously analysed for pp65Ag and PCR. Overall, 28/38 patients (74%) had active CMV infection within 100 days from SCT. In 16 patients, CMV was first detected by pp65 Ag alone; in 5 patients by both methods and in 6 by PCR assay alone; one patient had CMV biopsy-proven intestinal disease without pp65Ag and PCR assays positivity before CMV disease. Overall, three patients developed intestinal CMV disease (7.9%): one had negative both pp65Ag and PCR assays before CMV disease, one had disease and concomitant positivity of both methods, while in the remaining patient, only pp65Ag was positive before CMV disease.ConclusionPlasma PCR(COBAS AMPLICOR CMV MONITOR) and pp65Ag assays were effective in detecting CMV infection, however, discordance between both methods were frequently observed. Plasma PCR and pp65Ag assays may be complementary for diagnosis and management of CMV infection.

Evaluation of a score system for the severity and outcome of cytomegalovirus interstitial pneumonia in allogeneic bone marrow recipients

We verified whether a clinical score system developed for renal transplant patients predicts the severity and outcome of cytomegalovirus interstitial pneumonia (CMV IP) in allogeneic bone marrow (BMT) recipients. The score system was retrospectively applied to 20 patients at the estimated date of onset of IP and 10-14 days later. Seven patients received ganciclovir (GCV), seven received GCV plus intravenous immunoglobulin (i.v. Ig), and six received only supportive care. Nine out of 14 patients who received GCV with or without i.v. Ig survived the episode of IP (the median score of these patients at diagnosis of CMV IP was 5 (range 3-8)), while the remaining five patients died of respiratory failure during IP and at the diagnosis had a median score of 10 (range 9-11) (P=0.01). The six patients who received only supportive care survived for a median time of 21 days (range 10-24 days) from the estimated onset of CMV IP, and the median score at the diagnosis of IP was 10 (range 8-12). The overall survival correlates strongly with low initial severity of IP as measured by this score system: 11 out of 20 patients who died of respiratory failure during IP had at the estimated onset of IP a score >8, while of the nine patients who survived IP, eight had at the onset a score <7 and the remaining one a score of 8 (P=0.0007). The sensibility, specificity, predictive positive value and predictive negative value of the score system (with a threshold value of 8) to identify patients who survived IP was: 100%, 88%, 91% and 100%, respectively. The use of ganciclovir alone or in combination was the most important determinant of outcome. These data support the relevance of this score system with a threshold value of 8; if prospective and controlled studies confirm our observations, it would help physicians to identify BMT recipients during CMV IP with high vs. low risk of poor outcome.

Prospective cytomegalovirus monitoring during first-line chemotherapy in patients with acute myeloid leukemia

Little is known about the incidence and clinical impact of cytomegalovirus (CMV) infection in patients with acute myeloid leukemia at the time of diagnosis and during chemotherapy. The aims of the present study were to assess prospectively the incidence of active CMV infection in 69 consecutive patients with acute myeloid leukemia and to describe the outcomes of treatment. pp65 antigenemia was monitored at diagnosis, post‐induction and post‐consolidation chemotherapy, and whenever CMV reactivation was suspected. Patients with pp65 antigenemia received pre‐emptive anti‐CMV treatment. Fifty‐nine patients achieved complete remission. Baseline CMV serology results were available for 56 of the 59 patients: 52 patients (93%) were IgG positive. The overall incidence of pp65 antigenemia in patients in complete remission after chemotherapy was 35% (21/59): 9 patients after induction and 12 post‐consolidation. Sixteen of the 21 pp65‐positive patients received anti‐CMV treatment: 15 as pre‐emptive therapy and 1 for interstitial CMV pneumonitis. Five patients received no anti‐CMV treatment and did not develop CMV disease. Patients with pp65 antigenemia had more hospital admissions (2.57 vs. 2.16; P = 0.009), while patients with >10 pp65‐positive cells had more clinical complications (8/9 vs. 2/12; P = 0.002). In conclusion, patients with acute myeloid leukemia receiving chemotherapy should be monitored for active CMV infection. CMV reactivation in these patients was associated with an increased number of hospital admissions, and high levels of pp65 antigenemia were associated with more clinical complications. Controlled studies are needed to assess the relevance of pre‐emptive anti‐CMV therapy in patients with acute myeloid leukemia receiving chemotherapy. J. Med. Virol. 82: 1201–1207, 2010.

The immune reconstitution after an allogeneic stem cell transplant correlates with the risk of graft-versus-host disease and cytomegalovirus infection

Aim of the study was to correlate the clinical outcome of eighteen patients who have undergone an allogeneic stem cell transplant (SCT) with the concentration in the peripheral blood (PB) of lymphocyte subpopulations evaluated at 1 year from transplant. The occurrence of acute GVHD and CMV infection correlated with the concentration of Tregs in the PB; CMV infection also correlated with the content of NK cells. The obtained results document that the concentration of Tregs in the PB after an allogeneic SCT may protect from GVHD and from CMV infection; the potential anti-viral role of NK cells is confirmed.

Longitudinal analysis of human herpesvirus-8 DNA and antibodies in an Italian allogeneic stem cell transplant recipient

BACKGROUND Changes of HHV-8 antibody reactivity and intermittent detection of HHV-8 DNA have been observed in subjects with Kaposiss Sarcoma and/or HIV infection. Little is known about the longitudinal dynamics of HHV-8 DNA and antibody response in allografted stem cell transplant (SCT) patients without Kaposiss Sarcoma. OBJECTIVES To report the natural history of a HHV-8 seropositive patient with chronic lymphocytic leukemia who developed an active HHV-8 infection after SCT. STUDY DESIGN HHV-8 antibodies were measured by IFA and ELISA assays. HHV-8 DNA was detected by real-time PCR quantitative assay in serum and peripheral blood leukocytes (PBL). RESULTS Twenty-two out of 26 (85%) serum samples had detectable HHV-8 antibodies: 21/26 (80%) samples were positive by both IFA and ELISA assays, while 1 sample was ELISA positive-IFA negative. The remaining 4 samples (15%) were negative by both assays. Five out of 6 (83%) serum-PBL samples pairs had detectable HHV-8 DNA: a median of 934 genomes/ml (range 254-6316 genomes/ml) in the serum and a median of 10,000 genomes/10(5) (range 1472-93,460 gen/10(5)) in PBL. An active HHV-8 infection occurred early within the first 30 days after the transplant, extended up to day +180 and occurred without evidence of HHV-8-related neoplastic or non neoplastic diseases. CONCLUSIONS This study provides evidence that a patient infected with HHV-8 before SCT can be either intermittently HHV-8 DNA-positive and/or seropositive for HHV-8 after SCT. A high and persistent HHV-8 replication may be ongoing even in the absence of overt HHV-8-associated diseases.

Cytomegalovirus in Bone Marrow Cells Correlates with Cytomegalovirus in Peripheral Blood Leukocytes

ABSTRACT In allogeneic hematopoietic stem cell transplant recipients with bone marrow (BM) suppression, cytomegalovirus (CMV) pp65 antigenemia and DNA were detectable in peripheral blood leukocytes (PBL) and BM cells. A relationship between CMV infection of PBL and BM cells has been found.