COPD is associated with increased pro-inflammatory CD28null CD8 T and NKT-like cells in the small airways

We previously showed increased steroid resistant CD28null CD8+ senescent lymphocyte subsets in peripheral blood from COPD patients. These cells expressed decreased levels of the glucocorticoid receptor (GCR), suggesting their contribution to the steroid resistant property of these cells. COPD is a disease of the small airways. We therefore hypothesized that there would be a further increase in these steroid resistant lymphocytes in the lung, particularly in the small airways. We further hypothesized that the pro-inflammatory/cytotoxic potential of these cells could be negated using prednisolone with low-dose cyclosporin A. Blood, bronchoalveolar lavage, large proximal and small distal airway brushings were collected from 11 COPD patients and 10 healthy aged-matched controls. The cytotoxic mediator granzyme b, pro-inflammatory cytokines IFNγ/TNFα, and GCR were determined in lymphocytes subsets before and after their exposure to 1µM prednisolone and/or 2.5ng/mL cyclosporin A. Particularly in the small airways, COPD subjects showed an increased percentage of CD28null CD8 T-cells and NKT-like cells, with increased expression of granzyme b, IFNγ and TNFα and a loss of GCR, compared with controls. Significant negative correlations between small airway GCR expression and IFNγ/TNFα production by T and NKT-like cells (eg, T-cell IFNγ R= -.834, p=.031) and with FEV1 (R= -890) were shown. Cyclosporine A and prednisolone synergistically increased GCR expression and inhibited pro-inflammatory cytokine production by CD28null CD8- T and NKT-like cells. COPD is associated with increased pro-inflammatory CD28null CD8+ T and NKT-like cells in the small airways. Treatments that increase GCR in these lymphocyte subsets may improve efficacy of clinical treatment.

COPD is associated with increased pro-inflammatory CD28null CD8 T and NKT-like cells in the small airways

We previously showed increased steroid resistant CD28null CD8+ senescent lymphocyte subsets in peripheral blood from COPD patients. These cells expressed decreased levels of the glucocorticoid receptor (GCR), suggesting their contribution to the steroid resistant property of these cells. COPD is a disease of the small airways. We therefore hypothesized that there would be a further increase in these steroid resistant lymphocytes in the lung, particularly in the small airways. We further hypothesized that the pro-inflammatory/cytotoxic potential of these cells could be negated using prednisolone with low-dose cyclosporin A. Blood, bronchoalveolar lavage, large proximal and small distal airway brushings were collected from 11 COPD patients and 10 healthy aged-matched controls. The cytotoxic mediator granzyme b, pro-inflammatory cytokines IFNγ/TNFα, and GCR were determined in lymphocytes subsets before and after their exposure to 1µM prednisolone and/or 2.5ng/mL cyclosporin A. Particularly in the small airways, COPD subjects showed an increased percentage of CD28null CD8 T-cells and NKT-like cells, with increased expression of granzyme b, IFNγ and TNFα and a loss of GCR, compared with controls. Significant negative correlations between small airway GCR expression and IFNγ/TNFα production by T and NKT-like cells (eg, T-cell IFNγ R= -.834, p=.031) and with FEV (R= -890) were shown. Cyclosporine A and prednisolone synergistically increased GCR expression and inhibited pro-inflammatory cytokine production by CD28null CD8- T and NKT-like cells. COPD is associated with increased pro-inflammatory CD28null CD8+ T and NKT-like cells in the small airways. Treatments that increase GCR in these lymphocyte subsets may improve morbidity in COPD patients.

Natural killer cell deficiency experiences higher risk of sepsis after critical intracerebral hemorrhage

Background: Lymphopenia is common in intracerebral hemorrhage (ICH) and may predispose to severe infections such as sepsis. However, what specific kind of lymphocytes subsets decreases is still unclear. We investigated the impact of lymphocytes subsets on post-critical ICH infections and mortality. Methods: Consecutive ICH patients (admitted to a single center between January 2017 and January 2018) were prospectively assessed to evaluate the following main parameters: peripheral blood lymphocytes, infections, and clinical scores. Predicting factors of sepsis were measured using multivariate Logistic regressions analysis. A Kaplan–Meier survival curve was performed to compare the mortality between septic and nonseptic patients. Survival status was evaluated by multivariate Cox regression analysis. Results: In total, 112 critical ICH cases were enrolled including 29 septic patients. Total counts of lymphocytes decreased accordingly with reduced lymphocyte subsets, especially natural killer (NK) cells and CD8+T lymphocytes after ICH. Septic patients had a higher incidence of pneumonia, a longer length of stay, higher 90-day mortality, and worse long-term outcomes. Multivariate Logistic regression analysis showed venous catheterization, high APACHE-II score (>15), low GCS score (3–5), and NK cells percentages on admission were independently associated with ensuing sepsis. After sepsis, the percentages of CD4+T and NK cells percentages decreased, CD8+T cells increased followed by a significantly decreased CD4/CD8 ratio. Bloodstream infection alone directly affected the survival status of patients with sepsis. Conclusions: Critical ICH patients underwent immune dysfunction and NK cells deficiency could favor nosocomial threatening sepsis after ICH.

Immune Profiling Evaluation of Newly Diagnose Multiple Myeloma (NDMM) Transplant Eligible (TE) Patients Treated with Daratumumab, Cyclophosphamide, Thalidomide and Dexamethasone. Preliminary Results

Background: CD38-targeting antibody Daratumumab (Dara) has been demonstrating significant improvement in (MM) patient's survival. Cyclophosphamide (C), thalidomide (T) and dexamethasone (D) - (CTd) is one of the most used induction protocols worldwide and the MAX-Dara study was the first that combine Dara-CTd as induction for (NDMM) (TE) patients. We hypothesized that this new combo + autologous stem cell transplantation (ASCT) could affect the quantitative recovery of distinct lymphocytes subsets. Objective: Primary endpoint was to quantify lymphocytes subpopulations in (NDMM) (TE) patients at different treatment phases. Secondary endpoint was to evaluate B cells subsets at same times. Methods: Peripheral blood of 10 NDMM TE patients was collected at three different moments: at diagnose, after 4 induction cycles and after two consolidation cycles post- (ASCT). Dara-CTd protocol was for up to four 28-day induction cycles: C-500mg per oral (PO) d 1,8 and 15, T at 100-200mg PO d 1 to 28, Dex at 40mg PO d 1,8,15 and 22 and Dara 16mg/Kg/dose IV on d 1,8,15 and 22 during cycles 1 - 2 and every other week in cycles 3 - 4, followed by ASCT. Consolidation was started at D+30 after ASCT and all patients received up to four 28-day consolidation cycles: Dara 16mg/Kg and (D) at 40mg every other week, associated with T at 100mg PO d 1 - 28. Dara 16mg/Kg was used monthly as maintenance until progression or limiting toxicity. Flow cytometry was used to detect lymphocyte surface by CD3, CD4, CD5, CD8, CD16, CD19, CD20, CD38, CD45 and CD56 in the scatter plot. B cells were isolated and subpopulations (naïve B cells, class and non-class switched memory B cells, , IgD-CD27- memory B cells and plasma blasts) were detected by CD20, CD24, CD27, CD38, CD45 and IgD. Statistical analysis was performed using the SPSS® v25.0. Results: The median number of lymphocytes subsets at diagnosis were 1139 x 10³/μL for T cells, 155 x 10³/μL for B cells and 284 x 10³/μL for NK cells. After four cycles of Dara-CTD the median number of T, B and NK cells had dropped to 834, 7.5 and 8.0 x 10³/μL respectively (p<0.05). After two consolidation cycles post-ASCT, the T cells showed full reconstitution (1246 x 10³/μL) while B cells and NK cells had weakly reconstitution (20 x 10³/μL and 33 x 10³/μL, respectively). Regarding B cells subpopulations, the median B cell naïve numbers decreased from 32 x 10³/μL to 1 x 10³/μL (after 4 cycles), and recovery post-ASCT to 14 x 10³/μL. Class and non-class switched memory B cells numbers decreased after induction from 30 to 3.5 x 10³/μL and 37 to 2.0 x 10³/μL respectively. These subpopulations recovery after ASCT+ two consolidation cycles were not observed. Discussion: Different cells populations expresses CD38 antigen in their surface and depending on that, transitional lymphocytes counts reduction have been shown with different protocols using Dara. The present study confirmed that there is a decrease on total lymphocytes numbers after Dara- use. After two consolidation cycles post-ASCT, T cells counts had been recovered, while NK and B cells had a slightly recovery suggesting that Dara-CTD combination had a slighted negative impact in those lymphocytes' reconstitution. Concerning specifically B cells populations, we found that naive B cell was the first to showed faster recovery, although it was still below the reference range (33 - 259 x 10³/μL). Conclusion: This is the first study that reported lymphocyte profile with Dara plus CTD protocol. The preliminary data suggests that Dara-CTD reduces all lymphocytes populations after induction phase, but after ASCT followed by two consolidations cycles full reconstitution of T cells and slight recovery of B and NK cells was observed. Disclosures De Queiroz Crusoe: Janssen: Research Funding.

Immunohistochemistry of Lymphocytes Subsets in Subclinical Experimental Paratuberculosis in Goats

The present experiment was carried out to find out the lymphocytes subsets reactions in experimentally induced subclinical paratuberculosis in goats. Twelve goats of 8-12 weeks age were infected with 4.23 x 109 Mycobacterium avium paratuberculosis on 8 occasions. Seven goats were kept as in-contact controls and 4 as uninfected controls. Immunohitochemistry for detection of cellular reaction of CD2+, CD4+, CD8+, CD25+, MHC I and MHCII in the lymphocytes present in the intestine and lymph node revealed more reactive cells in the infected goats as compared with the in-contact and infected control goats.

Comparison of postoperative lymphocytes and interleukins between laparoscopy-assisted and open radical gastrectomy for early gastric cancer

Objective This study aimed to study the effects of laparoscopic-assisted radical gastrectomy (LAG) and open radical gastrectomy (OG) on immune function and inflammatory factors in patients with early gastric cancer. Methods Seventy-five patients with pT1N0M0 gastric cancer in Ren Ji Hospital from August 2017 to January 2018 were studied. Lymphocytes subsets and interleukins were compared preoperatively and on the third postoperative day (POD3) and seventh postoperative day (POD7). Results There were no significant differences in age, sex, body mass index, duration of the operation, estimated blood loss, total gastrectomy rate, postoperative first fluid diet, and the levels of preoperative lymphocytes subsets and interleukins between the two groups. The number of CD4+ T cells and the CD4+/CD8+ ratio in the LAG group were significantly higher than those in the OG group on POD3. However, the number of CD8+ T cells, and interleukin-6 and interleukin-8 levels in the LAG group were significantly lower than those in the OG group on POD3. Conclusions Laparoscopy can effectively reduce the levels of inflammatory factors and has less effect on the immune system than OG.