High responses were seen only in few animals of each haplotype and not in general. A polymorphism in the chicken CD8α gene was found in our experimental chicken lines, resulting in incapability to detect CD8α+ T cells using antibodies from the CT8 clone. Screening chickens with alternative find more antibodies showed that antibodies from the 2-398 clone were able to discriminate all CD8α+ cells from CD8α− cells, and consequently this antibody was used in a second vaccination experiment performed

with chickens of the haplotypes B13 and B130. This experiment showed a significant difference in antigen-specific proliferation of CD4+ T cells between the two lines, but not in CD8α+ T cell proliferation. Newcastle disease virus (NDV), an avian paramyxovirus, is a contagious virus causing Newcastle disease (ND). NDV is able to infect almost all species of birds, of which chickens are the most susceptible, in that some field strain infections of chickens have been shown to cause mortalities of 50% or more [1]. As the virus can be spread through domestic as well as wild birds, it is extremely important to

be able to convey Trichostatin A concentration efficient protection to chicken flocks. Vaccination is a widely used method in the control of viral diseases, including ND in the poultry production [2]. Improved knowledge of mechanisms involved in immunological protection in chickens is important in order to develop improved vaccines and optimize vaccine regimens. A standard evaluation of vaccination efficiency is measuring specific antibody titres to vaccines. However, it is also well known that the cellular immune Sitaxentan system is a key actor in vaccine-induced

antiviral immunity [3]. Thus, it was shown by Marino and Hanson [4] that ND-vaccinated bursectomized chickens that were unable to produce antibodies at a protective level still resisted challenge with ND. Furthermore, it has been shown that the level of humoral response after ND vaccination measured by HI titres does not correlate with the cellular response as measured by the under-agarose leucocyte-migration-inhibition technique [5]. In general, commercial ND vaccines are known to induce protective immunity with T cells playing a major role in clearance of the virus [6–8]. In relation to this, reliable techniques for a quantitative and qualitative evaluation of T cell responses upon vaccination are of great importance. One method to measure cellular responses in chickens is the antigen-specific T cell proliferation assay based on carboxyfluorescein diacetate-succinimidyl ester (CFSE) staining of peripheral blood mononuclear cells (PBMC) in order to trace the proliferating T cells. Measurement of proliferating cells as well as non-proliferating cells is subsequently taken by flow cytometry.

Although it remains poorly understood, the processing of Ag present on the whole parasite might not only follow a

different process but that process might be more efficient and the multiple Ag could persist as a depot, which in some instances is required for the maintenance of memory T cells. ABT 888 This study points the way for further analysis of the antigen or antigens that are recognized by the expanded CD8+ TEM cells. T cell clones can be derived from the livers of γ-spz-immune mice and used to screen P. berghei Ag. Further elucidation of the Ag recognized by the Vβ-bearing T cells should provide insights into the role of these cells in protective immunity to malaria and the mechanism by which predominant TCR are selected in the immune response to immunization with radiation-attenuated spz. We thank Isaac Chalom, Gina Donofrio, Caroline Ciuni and Zahra Parker for the provision of spz from hand-dissected mosquitoes, and expert technical assistance;

the Department of Entomology, WRAIR for infected mosquitoes; Dr Robert Schwenk for critical review of this manuscript; and the entire Krzych Laboratory for many useful discussions. This work is supported in part by a grant from the NIH AI46438 (UK) and by US Army Research and Materiel Command. The opinions expressed in this article are personal and are not to be construed selleck chemical as official positions of the United Fossariinae States Departments of Army, Defense, or Health and Human Services. “
“Biomedical Advanced Research and Development Authority, Washington, DC, USA Pfizer, San Diego, CA, USA The efficacy

of multi-agent DNA vaccines consisting of a truncated gene encoding Bacillus anthracis lethal factor (LFn) fused to either Yersinia pestis V antigen (V) or Y. pestis F1 was evaluated. A/J mice were immunized by gene gun and developed predominantly IgG1 responses that were fully protective against a lethal aerosolized B. anthracis spore challenge but required the presence of an additional DNA vaccine expressing anthrax protective antigen to boost survival against aerosolized Y. pestis. Immunization against Bacillus anthracis is dependent upon the production of an effective antibody response directed against the bacterium’s tripartite exotoxin comprised of protective antigen (PA, a nontoxic cell-binding element), lethal factor (LF, a metaloprotease), and edema factor (EF, a cyclic AMP modulator; Turnbull, 1991; Baillie & Read, 2001).

We therefore decided to undertake experimental work to characterize the nature of infiltrating lymphoid cells in order to gain insight into the mechanism of autoreactivity in vitiligo. Ten patients with active disseminated vitiligo who had been diagnosed within 3 months prior to their inclusion in the study (early disease) and 10 other patients who had been diagnosed more than 2 years previously (late disease) were enrolled into the study. None had ever received topical or systemic immunosuppressant therapy, and ‘early disease’ cases had had no therapy. check details All patients were aware of the risks and signed a Clinical Investigation Agreement to participate in the study. The study

protocol was approved by the Research and Ethics Committee of the Centro de Hematología y Medicina Interna de Puebla, Laboratorios Cínicos de Puebla, and Laboratorios Clínicos de Puebla de Bioequivalencia. Punch skin biopsies were obtained from all patients. All biopsies were fixed in 10% buffered

formaldehyde and paraffin-embedded by routine methods. Sections were then rehydrated by sequential immersion in xylene and decreasing water solutions of ethanol for immunochemical staining. Antibodies to CD1a, CD2, CD3, CD4, CD5, CD8, CD20, CD25, CD30, CD56, CD68 and CD79a were used to characterize the lymphoid infiltrates in all biopsies. Citrate pH6 buffer (Citrates®; Cell Marque, Rocklin, CA, USA) was used for Decitabine cell line antigenic recovery of CD3, an ethylenediamine tetraacetic acid (EDTA) Selleckchem Palbociclib pH8 buffer (Trilogy®; Cell Marque) for the recovery of CD1a, CD2, CD4, CD5, CD8, CD20, CD30 and CD56 and an EDTA pH6 buffer (Decleare®; Cell Marque) for CD25, CD68 and CD79a. Immunochemical staining was performed with the aid of an automated platform (Dakoautostainer plus®; Dako, Glostrup, Denmark), and an alkaline

phosphatase polymer (UltraVision Labeled Polimer®; Labvision) and Fast Red C were used to unravel the binding of the different antibodies [1, 27-29]. Different positive and negative control tissue samples were run simultaneously to ascertain the sensitivity and specificity of each antigen–antibody reaction in the system. Two independent and skilled professionals counted the proportions of cells expressing each of the antigens in each of the biopsies. At least 200 cells were counted to determine the percentages of infiltrating cells expressing each of the CD antigens that were searched. A statistical t-test for paired observations was used to compare the mean values of the percentages of the different cell types between early and late disease lesions infiltrates. The MedCalc® (Ostend, Belgium) software package was used for this purpose. Table 1 summarizes the mean values and standard deviations of such figures in both biopsies from early and late disease biopsies. Figure 1 depicts the main changes in the proportions of cell subsets in biopsies from patients with lesions less than 3 months old (Fig.

, 2004). Sequencing of a part of the 5′-UTR and the complete VP1 region was performed by a modification of previously described methods (el-Sageyer et al., 1998; Kilpatrick

et al., 1998; Liu et al., 2000; Szendrői et al., 2000). For sequencing of the 5′-UTR, cDNA was prepared by random hexamer-primed reverse transcription from virion RNA templates, followed by PCR amplification using primers ‘1’ (sense; position: 163–184 nt; 5′-CAAGCACTTCTGTTTCCCCGG-3′) and ‘3’ (antisense; position: 579–599 nt; 5′-ATTGTCACCATAAGCAGCCA-3′). VP1 sequences were amplified by PCR using primers Y7 (sense; position: 2395–2418 nt; 5′-GGGTTTGTGTCAGCCTGTAATGA-3′) and Q8 (antisense; position: 3475–3496 nt; 5′-AAGAGGTCTCTRTTCCACAT-3′), which also served as sequencing primers along

Raf inhibitor review with panPV1A (sense; position: 2935–2916 nt; 5′-TTIAIIGCRTGICCRTTRTT-3′) and panPV2S (antisense; position: 2895–2876 nt; 5′-CITAITCIMGITTYGAYATGT-3′) (Kilpatrick et al., 2004). All primer positions are relative to Poliovirus P3/Leon 12 a1b, GenBank accession Selleckchem Opaganib number X00925 (Stanway et al., 1983). PCR products were purified using PCR-Clean up-M Kit (Viogene, Sunnyvale, CA). The 5′-UTR and VP1 sequences described in this study were submitted to the GenBank library under accession numbers EU918372EU918382 and EU918384EU918390. In Hungary, mOPV was used for immunization campaigns from December 1959 up to 1992, after which tOPV was used. In 1960, a total

of 36 cases of VAPP following administration of mOPV were reported in Hungary: five cases were associated with poliovirus type 1 (two OPV recipients and three OPV contacts), Florfenicol one with type 2 (recipient), and eight with type 3 (five recipients and three contacts), specimens from 19 patients were negative for poliovirus, and three specimens were not tested. From 1961 to 1990, an additional 54 VAPP cases were reported: three cases were associated with type 1, seven with type 2, and 44 with type 3. In the original investigations, the best available methods were used for intratypic serodifferentiation (distinguishing vaccine-related poliovirus isolates from wild type), which tested for antigenic and phenotypic properties such as reproductive capacity of growth at 40 °C (rct40 marker), sensitivity of plaque formation to sulfated polysaccharides (d marker), and elution properties from Al(OH)3. Of the 52 cases of VAPP in Hungary associated with poliovirus type 3, 18 type 3 isolates from 15 children with VAPP [eight typ3 mOPV (mOPV3) recipients, four OPV contacts, and three with unknown OPV histories] were recovered from archival storage (Table 1). The 15 VAPP patients were geographically and temporally dispersed without any epidemiological associations. Characterization of the type 3 isolates from the VAPP patients using diagnostic RT-PCR confirmed that all 18 type 3 isolates were derived from the Sabin type 3 OPV strain, Leon 12 a1b.

(ii) Dynamic and less well-defined tolerance mechanisms that control autoreactive lymphocytes. More than 40 years ago,

Johnson & Setchel cannulated the rete testes of rams and collected testicular fluid for analysis, noting the very low concentrations of proteins, including immunoglobulins, when compared with serum and lymph.53 They proposed the presence of a blood–testis permeability barrier around the seminiferous tubules. Initially, this was felt to be at the level of the myoid cells surrounding the base of the tubule. Subsequently, Dym & Fawcett54 studied the tight junctions in the seminiferous epithelium and the peritubular contractile layer at high magnification in rats, investigating selleck kinase inhibitor the permeability of these junctions to lanthanum nitrate, a very small electron-opaque tracer used in testing the

patency of intracellular clefts. They demonstrated that the blood–testis barrier existed at the level of tight junctions between CCI-779 Sertoli cells, what created a basal compartment containing lanthanum, separated from an adluminal compartment that did not. Cell surface interactions in the seminiferous epithelium are very dynamic. This epithelium consists of columnar Sertoli cells, along whose surface different generations of germ cells progress toward the tubular lumen while undergoing spermatogenic differentiation.3,4 The Sertoli cell lateral membrane is involved in dynamic contact with the germ cells, as well as in connecting adjacent Sertoli cells to each other by a belt of occluding junctions that provide structural integrity to the BTB. The domains of the Sertoli cell involved in anchoring late spermatids as well as those involved in inter-Sertoli junctional contacts in which the Sertoli cell membrane is paralleled by a thick bundle of actin filaments,

the so-called ectoplasmic specialization (ES).4,10 The BTB physically divides the seminiferous epithelium into basal C1GALT1 and apical (or adluminal) compartments. Besides its function as an immunologic barrier to segregate post-meiotic germ cell antigens from the systemic circulation, it creates a microenvironment for germ cell development and confers cell polarity. During spermatogenesis, the BTB must physically disassemble permitting the passage of preleptotene and leptotene spermatocytes. Studies have shown that this dynamic process is regulated by transforming growth factor-beta 3 (TGF-beta-3) and tumor necrosis factor-alpha.55 Antisperm antibodies (ASA) are detected in approximately 70% of men who have undergone vasectomy.56 These antibodies have also been associated with obstructive azoospermia secondary to cystic fibrosis and with unilateral or bilateral congenital absence of the vans deferens.57 Autoimmunity to sperm can also occur following testicular trauma or following mumps orchitis, which may occur in post-pubertal men but is rare before puberty.

4C). A cross-sectional view of the intracellular compartment revealed that cells challenged with 50 ng of fluorescently labeled OVA showed large internalized aggregates, as confirmed by other researchers 23. In contrast, OVA-desensitized cells showed fewer and smaller fluorescent aggregates, and their visual appearance was similar to that of cells challenged at 4°C, in which crosslinked receptors were not internalized and appeared with small aggregates bound to the membrane. Since desensitized cells were hypo-responsive to further triggering doses of the same

antigen, we studied the response to Copanlisib datasheet a second triggering antigen. Cells sensitized with anti-DNP IgE and anti-OVA IgE were desensitized to OVA or to DNP and then challenged

with triggering doses of DNP-HSA or OVA, respectively. Cells desensitized to OVA responded (β-hexosaminidase release) to a triggering dose of 1 ng DNP-HSA, and cells desensitized to DNP responded to a triggering dose of 10 ng OVA (see Fig. 4D), indicating that mediators were not depleted after desensitization to one antigen and that desensitization disabled the specific response only to the desensitizing antigen. We then analyzed the specificity of the calcium responses. Cells desensitized Lumacaftor price to OVA had impaired calcium influx when triggered with 10 ng OVA, but the influx was restored by a triggering dose of 1 ng DNP-HSA (see Fig. 4E, red line), indicating that the calcium response 17-DMAG (Alvespimycin) HCl was compartmentalized by specific antigen. We then analyzed

specificity using confocal microscopy (see Fig. 4F). OVA-desensitized cells showed low internalization of labeled OVA antigen (green) as compared to the larger aggregates seen in OVA-activated cells. When OVA-desensitized cells were challenged with DNP-HSA (purple), the amount of internalization was comparable to that of DNP-HSA activated cells, indicating that desensitization left unaffected the specific mechanisms of cell activation and receptor internalization. Our understanding of IgE desensitizations has been limited by the paucity of in vitro mast cell models providing quantitative and qualitative insight into the early and late cell responses. Here, we present an in vitro 11-step model of mouse BMMC rapid IgE desensitization under physiologic calcium conditions and characterize its kinetics, effectiveness, antigen specificity and receptor internalization-associated events. We showed that desensitization is a dynamic process in which each step provides a platform for the next level of response reduction and that once desensitized, mast cells remain hypo-responsive to further antigen challenges.

e. small beads in the respiratory zone versus large beads in the conductive zone), as distinct sizes of bacteria-containing LEE011 order beads should induce distinct inflammatory responses. To investigate this hypothesis further we used an Encapsulation

Unit Nisco Var J30 (NISCO Engineering AG, Zurich, Switzerland), which enables production of distinct sizes of P. aeruginosa containing seaweed alginate beads, through variation of nozzle size, air pressure and alginate flow rate. The aim of the present study was to study the course of chronic P. aeruginosa in groups of mice challenged with large or small P. aeruginosa-containing beads. The alginate enters through a central needle. The exit nozzle, which is centrally in line with the axis of the needle, has been countersunk externally, leading to the aerodynamic effect so that the jet has a smaller diameter when passing the nozzle than before at the needle. The needle is enclosed in a pressure chamber with an exit through the orifice. The size of the drop is determined by the nozzle size, the

product flow rate and the pressure inside the chamber. The product flow rate is controlled by a syringe pump to be connected to the product nozzle. The pressure chamber is controlled by the pressure-controlling unit. The pressure set point is fixed with a potentiometer. The clinical isolate P. aeruginosa strain PAO579 Selleckchem Talazoparib was propagated from a freeze culture for 18 h and grown for 18 h at 37°C in Ox-broth (Statens Serum Institute, Copenhagen, Denmark). The overnight culture was centrifuged at 4°C and 4400 g and the pellet resuspended in 5 ml serum-bouillon (KMA Herlev Hospital, Herlev, Denmark). Protanal triclocarban LF 10/60 (FMC BioPolymer N-3002 Drammen, Norway) was

dissolved in 0·9% NaCl to an alginate concentration of 1% and sterile filtered. The bacterial culture was diluted 1:20 in seaweed alginate solution. The solution was transferred to a 10-ml syringe and placed into a syringe pump (Graseby 3100; Ardus Medical Inc., Watford, UK). The syringe pump controls and feeds the alginate to the Encapsulation Unit Nisco Var J30. The J30 uses a pressure chamber containing a needle that controls the flow of alginate. The pressure chamber is controlled by the pressure controlling unit. The pressure set point is fixed with a potentiometer. The J30 unit is equipped with two connections, one for the alginate and one for the airflow that drives the alginate from the needle through the exit orifice into a gelleting bath (0·1 M, pH 7·0 Tris HCL buffer containing 0·1 M CaCl2). A magnetic stirrer (IKA RCT Basic; IKA®-Werke GmbH & Co. KG, Staufen, Germany) is placed underneath the gelling bath to prevent the beads from sticking together during gelling. The distance between nozzle and gelling bath of 11 cm and 280 rpm magnetic stirrer were kept constant. Five ml of alginate beads were made.

8,12,13 There are six alpha defensins: human neutrophil peptide (HNP)1–4 and human defensin (HD) 5 and 6. HNPs 1–3 share a high degree of homology with only the

amino terminal amino acid differing between them. Alpha defensins are synthesized as pre-prodefensins that are cleaved by proteases Selleck Selumetinib to create an active peptide which displays antibacterial activity against Gram-positive and Gram-negative bacteria, fungi, and yeast; and antiviral effects against HIV-1, HSV-1, and HSV-2.12 Intriguingly, however, HD5 and HD6 enhance HIV replication by themselves as well as in the presence of gonorrheal infection.20 However, the exact mechanism of infection remains to be determined. Beta defensins HBD1–6 are structurally similar to alpha defensins and have broad inhibitory activity against a range of pathogens including HIV-1.12 Genome scans have revealed at least 28 putative human beta defensins; though, only six have been discovered, of which four are present in the FRT.8,12,13 HBD1–3 have direct and indirect anti-HIV-1 activity.21,22 Similar to other antimicrobials, they interact directly with the viral envelope.12,21 Furthermore, find more they act upon target cell populations to decrease levels of the HIV-1 CXCR4 co-receptor as well as inhibit

the early steps of viral replication.21–23 Cathelicidins are a family of cationic antimicrobial peptides of which only one is found in humans, cathelicidin (hCAP-18/LL-37).24 LL-37 is present in the FRT and is composed of three domains: a signal peptide region, an N-terminal cathelin-like domain, and a C-terminal antimicrobial domain.9,24 The mature

peptide LL-37 is generated from hCAP-18 by protease cleavage, is broadly antibacterial, and inhibits HIV-1 replication Dimethyl sulfoxide in vitro independently of changes in HIV-1 co-receptor expression. Intriguingly, the cathelin-domain also has antibacterial activity but no disclosed anti-HIV-1 activity.5,25 Uniquely, hCAP-18 is cleaved to form ALL-38 by gastricsin, a protease present in seminal fluid that is reaction dependent on low pH found in the vagina.26 ALL-38 has a similar antibacterial profile to LL-37, but its anti-HIV activity is unknown. This remarkable mechanism for antimicrobial activation highlights the importance of male sexual fluids in modulating the protective response in the FRT.9,13 Secretory leukocyte protease inhibitor and Elafin, located together on chromosome 20, are members of whey acidic protein (WAP) family that possess a conserved whey four disulfide core domain (WFDC).27,28 The pair are endogenous protease inhibitors involved in the control of inflammatory responses and tissue remodeling.27,28 Unlike SLPI, Elafin is relatively restricted in its target population acting mainly on neutrophil and pancreatic elastase and neutrophil proteinase 3. Both proteins also demonstrate anti-HIV-1 activity that is independent of their protease inhibitor function.

Systemic autoimmune diseases can be modeled in transgenic mice harboring defects in DC apoptosis 10 but not in mice with apoptosis defects in T and B cells 11–13. Our study shows that in addition to the dogma of DC apoptosis as a mechanism to eliminate activated DC to prevent hyperactivation of the immune response, DC apoptosis also plays an

active role in induction and maintenance of tolerance through induction of Treg, whereby defects in DC apoptosis may trigger autoimmunity. High levels of spontaneous DC apoptosis have also been observed in breast cancer patients, with its significance being unclear 15, 16. Our study indicates that DC apoptosis in cancer patients may play a role in suppressing immune responses against the tumor by inducing immunosuppression and tolerance. Therefore, prevention Metformin research buy of DC apoptosis may enhance the therapeutic

effects of chemotherapy in tumor CH5424802 datasheet eradication 15, 16. Our findings may also represent a therapeutic approach in the prevention of unwanted immune responses in autoimmune diseases and transplantation along with inhibition of DC apoptosis to assist in tumor eradication. C57BL/6 mice were purchased from Charles River Laboratories (St. Constant, QC) and maintained as per guidelines of SickKids animal facilities. All the animal studies were reviewed and approved by the SickKids Institutional Committee for humane use of laboratory animals. OT-II mice were purchased from Jackson Laboratories (Bar Harbor, ME). The following antibodies were purchased from eBioscience (San Diego, CA): CD11c PE, CD86 PE, CD80 PE, MHC II PE, IL-10 Alexa647, IL-12 APC, IL-17 PE, Foxp3 PE along with neutralizing

IL-4 and IFN-γ Ab, and the following from BD Biosciences (Mississauga, ON): CD11c-FITC, CD4-FITC and CD3-PE. Anti-TGF-β neutralizing Ab (MAB1835) was obtained from R&D Systems (Minneapolis, MN). Isotype control IgG were obtained from eBioscience and/or PLEKHM2 Serotec (Raleigh, NC). CFSE was obtained from Molecular Probes (Burlington, ON); BrdU, OVA, cytochalasin D, rapamycin and PI were obtained from Sigma-Aldrich (Oakville, ON). GM-CSF was obtained from R&D Systems. IL-6 and TGF-β were obtained from Peprotech (Rocky Hill, NJ). Bone marrow cells were isolated from tibia and femurs of adult mice and cultured in the presence of GM-CSF for 7 days as described previously 34. DC were harvested and stained with 1 μM CFSE as described previously 35. Naïve CD4+CD25–CD62L+ T cells were isolated from spleens of mice using CD4+CD62L+ naïve T-cell isolation kit in conjunction with MACS columns from Miltenyi Biotec (Auburn, CA), following the manufacturer’s instructions. DC were cultured on a six-well dish and irradiated for 2 min with a UV transilluminator, with a peak intensity of 9000 mW/cm2 at the filter surface and a peak emission of 313 nm.

Importantly, these frequencies are very similar to those of humans [7]. Since there was some degree of nonspecific staining with the vehicle-CD1d dimers, a control staining with vehicle-CD1d dimers was always carried out in parallel to each α-GalCer-dimer staining. The calculation of the final frequencies, including the phenotypic characterization, was carried

out by subtracting the event numbers obtained with the vehicle-CD1d dimers from the event numbers obtained with α-GalCer-CD1d dimers. Results obtained with groups of three to ten individual animals are summarized in Table 1 of the Supporting Information while Figure 2 illustrates how these frequencies were calculated. In the spleen, a higher binding of the vehicle-loaded control dimers and the secondary reagent used to visualize Estrogen antagonist them was observed compared with that of the liver (Fig. 2A and Supporting Information Fig. 2). This nonspecific staining may have obscured specific binding of α-GalCer-CD1d-dimers to splenocytes and therefore a cautious interpretation of the final frequencies calculated for the spleen is warranted. A reliable phenotypic characterization of iNKT cells was only possible in the liver, but not for the extremely small

numbers of splenic iNKT cells. Most rat iNKT cells are DN or CD4+ (Fig. 2A and Supporting Information Table 1) and, similar to their mouse counterparts [17], rat iNKT cells express lower levels of CD4 than non-iNKT CD4+ T cells (Fig. 2C). Similar to humans, but in contrast to mice [1, 6], a fraction of rat iNKT cells were found to express https://www.selleckchem.com/products/BEZ235.html CD8α+ (Fig. 2C). In contrast, almost no CD8β+ iNKT cells were detected (Fig. 2A and Supporting Information Table 1). As shown in Figure 2, the majority of rat iNKT cells expressed NKR-P1A/B at intermediate levels, detected by the mAb 10/78 [18], but only a small fraction of all NKR-P1A/B+ T cells are iNKT cells (Supporting Information Table 1). It is important to highlight this finding since coexpression of NKR-P1A/B and TCR are used as surrogate markers for iNKT cells in the rat [19-21]. TCR usage among

iNKT cells was analyzed Anidulafungin (LY303366) with the mAbs R78 and HIS42, which in F344 rats bind, respectively, to TCRs containing BV8S4A2 (a homologue to mouse BV8S2) and BV16 [10]. About half of iNKT cells were R78 (BV8S4A2)-positive while almost no BV16+ iNKT cells were detected (Fig. 2C and data not shown for BV16). Recently, the transcription factor PLZF was found to be expressed at high levels by iNKT cells and to control the development of their innate-like features [22, 23]. As shown in Fig. 2(D), most iNKT cells, defined by concomitant staining with α-GalCer-CD1d dimers and anti-TCRβ mAb (as shown in Fig. 2A and C), were stained by an anti-PLZF mAb. Moreover, almost all PLZF+ αβ T cells were also stained by α-GalCer-CD1d dimers but not by vehicle-loaded CD1d dimers (Fig. 2E). Some TCRβ-negative cells were also stained with the anti-PLZF mAb (Fig. 2E).