A plasmablast biomarker for nonresponse to antibody therapy to CD20 in rheumatoid arthritis

An important goal for personalized health care is the identification of biomarkers that predict the likelihood of treatment responses. Here, we tested the hypothesis that quantitative mRNA assays for B lineage cells in blood could serve as baseline predictors of therapeutic response to B cell depletion therapy in subjects with rheumatoid arthritis (RA). In samples from the REFLEX trial of rituximab in inadequate responders to antibodies to tumor necrosis factor-α, a 25% subgroup of treated subjects with elevated baseline mRNA levels of IgJ, a marker for antibody-secreting plasmablasts, showed reduced clinical response rates. There were no significant efficacy differences in the placebo arm subjects stratified by this marker. Prospective testing of the IgJ biomarker in the DANCER and SERENE rituximab clinical trial cohorts and the SCRIPT ocrelizumab cohort confirmed the utility of this marker to predict nonresponse to anti-CD20 therapy. A combination mRNA biomarker, IgJhiFCRL5lo, showed improved test performance over IgJhi alone. This study demonstrates that baseline blood levels of molecular markers for late-stage B lineage plasmablasts identify a ~20% subgroup of active RA subjects who are unlikely to gain substantial clinical benefit from anti-CD20 B cell depletion therapy.     

Combined natural killer T-cell based immunotherapy eradicates established tumors in mice

A rational monoclonal antibody (mAb)-based antitumor therapy approach has previously been shown to eradicate various established experimental and carcinogen-induced tumors in a majority of mice. This therapy comprised an agonistic mAb reactive with tumor necrosis factor-related apoptosis-inducing ligand receptor (DR5), expressed by tumor cells, an agonistic anti-CD40 mAb to mature dendritic cells, and an agonistic anti-4-1BB mAb to costimulate CD8(+) T cells. Because agonists of CD40 have been toxic in patients, we were interested in substituting anti-CD40 mAb with other dendritic cell-maturing agents, such as glycolipid ligands recognized by invariant natural killer T (iNKT) cells. Here, we show that CD1d-restricted glycolipid ligands for iNKT cells effectively substitute for anti-CD40 mAb and reject established experimental mouse breast and renal tumors when used in combination with anti-DR5 and anti-4-1BB mAbs (termed "NKTMab" therapy). NKTMab therapy-induced tumor rejection was dependent on CD4(+) and CD8(+) T cells, NKT cells, and the cytokine IFN-gamma. NKTMab therapy containing either alpha-galactosylceramide (alpha-GC) or alpha-C-galactosylceramide (alpha-c-GC) at high concentrations induced similar rates of tumor rejection in mice; however, toxicity was observed at the highest doses of alpha-GC (>250 ng/injection), limiting the use of this glycolipid. By contrast, even very low doses of alpha-c-GC (25 ng/injection) retained considerable antitumor activity when used in combination with anti-DR5/anti-4-1BB, and thus, alpha-c-GC showed a considerably greater therapeutic index. In summary, sequential tumor cell apoptosis and amplification of dendritic cell function by NKT cell agonists represents an exciting and novel approach for cancer treatment.     

Renal transplantation in patients with Balkan endemic nephropathy

BACKGROUND: Balkan endemic nephropathy (BEN) is a chronic tubulointerstitial disease prevalent in Croatia, Romania, Bulgaria, Bosnia and Herzegovina, and Serbia. In addition to renal disease, an increased incidence of upper urothelial carcinomas (UUCs) has been observed in the foci of BEN. Carcinoma may occur alone or in combination with BEN. Immunosuppression is associated with an increased risk for development of different malignancies. There are no data in the literature about the outcome of patients with BEN after transplantation. METHODS: We performed a retrospective evaluation of the database and review of the charts and pathology reports of 601 renal transplant recipients treated at our institution. RESULTS: From January 1995 to December 2004, kidney transplantations were performed in nine patients with BEN. One-year graft survival was 100%. A man, who was transplanted in 1997 died 2 years after transplantation with a functioning graft due to disseminated cancer from the pelvis of his own kidney. A female patient developed UCC 2 years after transplantation. They were both treated with a bolus of methylprednisolone before transplantation, because of four HLA-mismatches. A male patient developed UCC in the native and transplanted kidneys. He underwent a native nephroureterectomy with partial nephroureterectomy of transplanted kidney. His graft function was preserved with decreased immunosuppression. Three years later a urinary bladder carcinoma was discovered on a regularly performed multislice computed tomography. One patient developed a skin malignancy. Other patients have had uneventful posttransplantation courses with excellent graft function. Thus, 33.3% of patients with BEN developed UUC, compared with a 0.67% prevalence of urinary tract tumors among transplanted patients with other causes of end-stage renal disease. CONCLUSION: Patients with BEN are at increased risk for the development of UCC after transplantation. Regular screening for early detection of malignancy is mandatory. Longer follow-up and results from other transplant centers are needed to further investigate the relationship between BEN and UCC after renal transplantation.     

Recognition of pollen-derived phosphatidyl-ethanolamine by human CD1d-restricted gamma delta T cells

BACKGROUND: Evidences from mice and human beings indicate that gammadelta T cells could be relevant in recognition of stress-induced self and/or yet unidentified inhaled foreign antigens. Their specificity differs from classic MHC-restricted alphabeta T cells and involves the immunoglobulin-like structure of the gammadelta T-cell receptor with the recognition of small organic molecules, alkylamines, and self lipid compounds presented by CD1+ dendritic cells. OBJECTIVE: Because CD1 receptors are mainly devoted to lipid antigen presentation, we sought to determine whether exogenous pollen membrane lipids may act as allergens for CD1-restricted gammadelta T cells. METHODS: Peripheral blood and nasal mucosa-associated gammadelta T cells were cloned from normal controls and cypress-sensitive subjects and tested for their antigen specificity and CD1-restriction with phospholipids extracted from tree pollen grains, as well with other natural or synthetic compounds. Phospholipid reactivity of cloned gammadelta T cells was measured by mean of proliferative response and cytokine release as well as by testing their helper activity on IgE production in vitro and in vivo. RESULTS: Cloned gammadelta T lymphocytes from subjects with allergy, but not normal controls, were found to recognize pollen-derived phosphatidyl-ethanolamine (PE) in a CD1d-restricted fashion. Only 16:0/18:2 and 18:2/18:2 PE were stimulatory, whereas no response was recorded for disaturated PE, phosphatidylcholine, neutral lipids, or protein extract. Proliferating clones secreted both T(H)1-type and T(H)2-type cytokines and drove IgE production in vitro and in vivo. CONCLUSION: CD1d-restricted gammadelta T cells specific for phospholipids can represent a key mucosal regulatory subset for the control of early host reactivity against tree pollens. CLINICAL IMPLICATIONS: By knowing how lipid allergen constituents interact with mucosal immune system, we can expand our possibilities in diagnostic and therapeutic interventions.     

Minilaparoscopy and open laparoscopy: prevention of laparoscopic complications

BACKGROUND: The study aimed to compare laparoscopy, open-laparoscopy and mini-laparoscopy and to correlate the results of each technique with the respective percentages of laparotomic conversion. METHODS: A total of 101 laparoscopies were performed between November 1997 and April 1999: 18.8% were diagnostic and 81.2% operative. The latter included 54 traditional laparoscopies (65.9%), 18 open-laparoscopies (21.9%) and 10 minilaparoscopies (12.2%). RESULTS: Laparotomic conversion was required in 5.5% of laparoscopies. No laparotomic conversion was necessary for the open-laparoscoples and for mini-laparoscopies. CONCLUSIONS: The possibility of resorting to open-laparoscopy and mini-laparoscopy may represent a valid tool when the patient has previously undergone laparoscopic or laparotomic surgery, with the supposition of pelvo-abdominal adherences that would increase the risk of traditional laparoscopy.     

Role of capacitation in intrauterine insemination as a treatment of male infertility

BACKGROUND: Although FIVET and ICSI efficacy and efficiency are continuously increasing, intrauterine insemination (IUI) is a very used technique for many different types of sterility. It is also used in male sterility for its cost/benefit positive rate. Pregnancies frequency obtained after IUI in male factor infertility cases, with or without ovarian stimulation, shows the value of this work. METHODS: We evaluated 149 insemination cycles in 34 couples with a male infertility diagnosis confirmed after at least 2 semen analysis, according to OMS criteria. All the 34 patients had at least one Fallopian tube open and some spontaneous ovulatory cycle to enter in this study. The patients without biochemical pregnancy signs after 6 intrauterine insemination cycles were induced to multiple ovulations through oral administration of 50 mg of Clomifene citrate from the 5th to the 9th cycle-day or through intramuscular administration of 75 I.U. of FSH from the 5th cycle-day. A luteal support with 200 mg of vaginal progesterone, 2 times for day, was reserved for all the patients with endometrium <8 mm or with low progesterone serum levels (<20 nmol/l). RESULTS: All patients made 149 IUI cycles, with a total medium pregnancy rate for cycle of 6.0% and for patient of 26.4%. We didn't observe statistically significant differences related to spermatozoa total number for ml in native sperm, in terms of pregnancy rate in men with 100-150 millions of spermatozoa. We observed a lower pregnancy rate in men with less than 5 millions moving spermatozoa. Under 1 million of total spermatozoa, pregnancy rate is very low (2.3%): it is established for higher values, obtaining the best results between 10 and 20 millions of total spermatozoa (10.5%) and a little reduction over 20 millions (8.3%). CONCLUSIONS: The evaluation of spermatozoa total number and also their motility does not present any prognostical significance as to the pregnancy rate in intrauterine insemination for male factor infertility.     

Human CD1b and CD1c isoforms survey different intracellular compartments for the presentation of microbial lipid antigens

CD1b and CD1c are antigen-presenting molecules that mediate recognition of bacterial lipids by T cells, but it is currently not known whether these two molecules are redundant or are specialized to perform different immunological functions. Here, we show that the distribution of CD1c in human dendritic cells was characterized by a high ratio of cell surface to intracellular molecules, whereas CD1b showed a reciprocal pattern of distribution. In contrast to the accumulation of CD1b in lysosomal major histocompatibility complex class II compartments, intracellular CD1c molecules accumulated in other endocytic compartments, most likely early and late endosomes. Deletion of the cytoplasmic tail of CD1c, containing a tyrosine-based internalization motif, abolished most of its intracellular localization. Functional studies using T cells specific for defined lipid antigens revealed that in contrast to CD1b-mediated antigen presentation, antigen presentation by CD1c was resistant to drugs inhibiting endosomal acidification and was independent of endosomal localization of CD1c. Taken together, these results support the hypothesis that CD1b and CD1c are specialized to survey the lipid content of different intracellular compartments.