Characterization of 25 monoclonal antibodies to factor VIII-von Willebrand factor: relationship between ristocetin-induced platelet aggregation and platelet adherence to subendothelium

We have studied the role of factor VIII-von Willebrand factor (FVIII- vWF) in both platelet adherence to subendothelium and ristocetin- induced platelet aggregation using monoclonal antibodies to human FVIII- vWF. Twenty-five monoclonal antibodies were obtained, two of which were directed to the factor VIII moiety of FVIII-vWF; one of these two completely inhibited the procoagulant activity (FVIII:C). The remaining 23 monoclonal antibodies were directed to the von Willebrand factor moiety of FVIII-vWF. The ability of the latter monoclonal antibodies to inhibit platelet adherence to arterial subendothelium was investigated with a perfusion model. According to the number of platelets adhering to the subendothelium, three groups of monoclonal antibodies could be discerned: (A) antibodies not affecting platelet adherence; (B) antibodies that inhibited platelet adherence to the level as observed when von Willebrand's disease plasma was tested; and (C) antibodies that completely inhibited both platelet adherence to subendothelium and ristocetin-induced platelet aggregation. The two antibodies present in group C competed for the same or closely related epitope(s) present on FVIII-vWF. These results demonstrate that a domain is present on the FVIII-vWF molecule that is associated both with ristocetin-induced aggregation and with the ability of FVIII-vWF to support platelet adherence to the subendothelium. Based on these observations, it is concluded that ristocetin-induced binding of FVIII-vWF to platelets reflects, at least in part, a physiologic mechanism regulating the function of FVIII-vWF in primary hemostasis.     

Potential capsule switching from serogroup Y to B: The characterization of three such Neisseria meningitidis isolates causing invasive meningococcal disease in Canada

Three group B Neisseria meningitidis isolates, recovered from meningococcal disease cases in Canada and typed as B:2c:P1.5, were characterized. Multilocus sequence typing showed that all three isolates were related because of an identical sequence type (ST) 573. Isolates typed as 2c:P1.5 are common in serogroup Y meningococci but rare in isolates from serogroups B or C. Although no serogroup Y isolates have been typed as ST-573, eight isolates showed five to six housekeeping gene alleles that were identical to that of ST-573. This suggested that the B:2c:P1.5 isolates may have originated from serogroup Y organisms, possibly by capsule switching.Key Words: Capsule switching, Neisseria meningitidis, Serogroup YNeisseria meningitidis is a significant pathogen that causes invasive meningococcal disease (IMD). The average case fatality rate of 9% to 12% remains high despite the availability of effective antibiotics and vaccines (1). Laboratory study and surveillance of N meningitidis involves the characterization of a number of surface markers of the bacterium, including its capsule and outer membrane proteins (OMPs). Most epidemiological studies of meningococcal disease rely on differentiating meningococcal isolates based on their serogroup, serotype and serosubtype. Serogrouping is determined by the demonstration of serologically distinct epitopes present on chemically and structurally different capsules. Serotyping and serosubtyping rely on the detection of distinct epitopes present on three of five different classes of OMPs of N meningitidis. Serotyping epitopes are found on the class 2 or class 3 OMP (also called PorB) of N meningitidis; these OMPs are expressed in a mutually exclusively manner (ie, a strain will only express either a class 2 or class 3 OMP but not both). Serosubtyping epitopes are present on the class 1 OMP (also called PorA). Based on this nomenclature scheme, a strain can therefore be characterized by its antigenic formula; for example, B:15:P1.7,16 refers to serogroup B, serotype 15 and serosubtype P1.7,16.One of the most important virulence factors of meningococci is the capsular polysaccharide antigen, which is also the basis for serogrouping and is the target antigen for the currently licensed vaccines against A, C, Y and W135 organisms. Of the 13 known serogroups, five (serogroups A, B, C, Y and W135) are responsible for most of the meningococcal disease worldwide (2). In North America, most endemic and epidemic strains belong to serogroups B, C, Y and W135 (3,4). Capsules of serogroups B, C, Y and W135 meningococci contain sialic acid, either as a homopolymer of sialic acids assembled by alpha-2,8 linkages (serogroup B) or alpha-2,9 linkages (serogroup C), or as a heteropolymer of sialic acids with glucose (serogroup Y) or galactose (serogroup W135). Besides demonstrating structural similarities, these four serogroups of meningococci also have very similar capsule polysaccharide synthesis (cps) gene loci (5). Because of this similarity, capsule switching has been demonstrated in vivo and in vitro by specific gene replacement within the cps loci between different serogroups. To date, a number of IMD cases have been described in the literature to be caused by organisms in which capsule switching between serogroup B and C meningococci occurred (6-8).In the present paper, the authors describe three unusual serogroup B meningococci isolated from separate IMD cases in Nanaimo, British Columbia, that presented with the OMP antigens 2c:P1.5, characteristic of serogroup Y strains found in Canada (4). This antigenic profile prompted the authors to examine the relationship of these three serogroup B strains with antigenically similar serogroup Y organisms isolated in Canada. The authors describe the characterization of these antigenically similar isolates and postulate that the B:2c:P1.5 isolates arose by capsule switching from serogroup Y organisms.     

Children and adolescents with marginal zone lymphoma have an excellent prognosis with limited chemotherapy or a watch‐and‐wait strategy after complete resection

Data on management of pediatric marginal zone lymphoma (MZL) are scarce. This retrospective study assessed characteristics and outcome in 66 patients who were <18 years old. Forty‐four (67%) had an extranodal MZL (EMZL), 21 (32%) a nodal MZL (NMZL), and one patient a splenic MZL. Thirty‐three patients (50%) received a variable combination of adjuvant chemotherapy/immunotherapy/radiotherapy, while the remainder, including 20 of 21 with NMZL, entered an active observation period. Overall survival was excellent (98 ± 2%), although 11 patients relapsed (17%; NMZL, n = 1; EMZL, n = 10), seven after any therapy and four after complete resection only. In conclusion, outcome of NZML, in particular, seems to be excellent after (in)complete resection and observation only.     

T cell immune profiling of respiratory syncytial virus for the development of a targeted immunotherapy

Respiratory syncytial virus (RSV)-associated viral infections are a major public health problem affecting the immunologically naïve/compromised populations. Given the RSV-associated morbidity and the limited treatment options, we sought to characterize the cellular immune response to RSV to develop a targeted T cell therapy for off-the-shelf administration to immunocompromised individuals. Here we report on the immunological profiling, as well as manufacturing, characterization and antiviral properties of these RSV-targeted T cells. A randomized, phase 1/2 clinical trial evaluating their safety and activity in haematopoietic stem cell transplant recipients as an off-the-shelf multi-respiratory virus-directed product is currently underway (NCT04933968, https://clinicaltrials.gov ).