69 Indeed, the Ras-MEK-MAPK, Rac1, and PI3K-Akt-mTOR signaling pathways involved in JSRV-induced cell transformation are important regulators of trophoblast growth and differentiation in human and rodent placentae.69 ERVs are present in the genomes of all vertebrates2 and can be used as DNA fossils to unravel virus–host coevolution over millions of years.8 The domestic sheep constitutes a powerful model to study the biological significance of ERVs given the contemporary presence in this animal species of a pathogenic exogenous retrovirus (JSRV) and the biologically active enJSRVs. Indeed, the study of enJSRVs provided the first in vivo evidence https://www.selleckchem.com/screening/mapk-library.html of a physiological role for ERVs in conceptus

and placental development.66 Collective evidence from studies of primates, rodents, rabbits, and sheep supports the idea that independent ERVs influenced mammalian evolution and were positively selected for a convergent physiological role in placental morphogenesis. Finally, it is likely that ERVs have other biological roles in reproduction including protection of the host reproductive tract from infectious and pathogenic exogenous retroviruses as well as fetomaternal tolerance. We are grateful to the members of the Laboratory for Uterine

Biology and Pregnancy of Texas A&M University and the Laboratory of Viral Pathogenesis of the University of Glasgow Faculty of Veterinary Medicine for stimulating discussions. Work in the laboratory of the authors is supported by NIH grant HD052745, a program grant of the Wellcome Trust and by a Strategic Research Developmental Grant by the Scottish Sirolimus Cepharanthine Funding Council. “
“Fibroblast heterogeneity has been recognized for decades, but the basis for multiple phenotypes among these cells has been investigated only recently. More than 15 years ago, Bucalla and his colleagues described for the first time a population of fibroblast-like cells among circulating mononuclear blood cells. Subsequently these mesenchymal cells, termed fibrocytes, have been characterized and found

to participate in normal and pathological tissue remodelling. In this review, I have attempted to present the evidence generated thus far suggesting that fibrocytes are participants in autoimmune diseases where tissues are injured and undergo remodelling. Aspects of their phenotype suggest that they are well suited to help orchestrate immune responses through mononuclear cell recruitment and their ability to produce inflammatory mediators and extracellular matrix molecules. These attributes also raise the possibility that they might be useful targets against which therapeutic agents might be aimed. Fibroblast heterogeneity has been appreciated for several decades but its biological significance and the basis for cellular diversity remain uncertain. The question of why fibroblasts from distant anatomical regions should exhibit phenotypic divergence is unanswered.

Under Th17 conditions, the binding of Mel-18 at the Ifng promoter was much lower than at the Il17a promoter (Fig. 4H). We did not notice changes in the binding activity of Mel-18 at Hoxa7 promoter in the presence or absence of Th17 polarizing cytokines (Fig. 4G). As with Mel-18, there were no significant changes BMS-777607 in the expression levels of the mRNA or protein of Ezh2 if the restimulation was either in the presence of Th17 conditions or IL-12 (Fig. 5A and B). But in contrast to Mel-18, the binding activity of Ezh2 at the Il17a promoter was not decreased without cytokines (Fig. 5C). The binding of Ezh2 at the Rorc,

Ifng, Tbx21 and Hoxa7 promoters was also not significantly altered between the different conditions (Fig. 5D–G). Ezh2 was associated more strongly with the Il17a promoter than with the Ifng promoter (Fig. 5H), but the differences were smaller in comparison to the differential binding activity of Mel-18 at these promoters (Fig. 4H). To determine whether the signaling pathways downstream to TGF-β were sufficient to maintain the high level of the binding activity of Mel-18 at the Il17a promoter, Th17 cells were restimulated

without cytokines or in the presence of either TGF-β alone or combination of TGF-β, IL-6 and IL-23 (Fig. 6A). The binding of Mel-18 was only modestly decreased when the restimulation was in the presence see more of TGF-β alone than with the cytokine combination. When the cells were restimulated without cytokines, the binding was further reduced almost to the level of unstimulated cells (resting). These results show that TGF-β is required for the maintenance of the binding activity of Mel-18 at the Il17a promoter

beyond the early TCR-dependent stage. Nevertheless, the presence of TGF-β in the absence of TCR stimulation, as in the resting conditions, is insufficient to induce the binding activity of Mel-18 at the Il17a heptaminol promoter. The binding activity of RORγt was correlated with this of Mel-18; RORγt was associated with the Il17a promoter when the Th17 cells were restimulated for 18 h in the presence of the Th17 polarizing cytokines but not in their absence (Fig. 6B). The decrease in the binding activity of RORγt may reflect the reduced expression of Rorc mRNA following restimulation without TGF-β (Fig. 3A). However, we did not recognize substantial changes in the expression levels of RORγt protein at this time point (Fig. 6C). Therefore, as early as 18 h following restimulation the recruitment of RORγt, and not its expression, is regulated by the polarizing cytokines.

These cross-reactive T cells were found to be subdominant during the primary response, and the sequence of infection influenced the

selleck compound hierarchy of these subdominant cross-reactive T cells after secondary heterologous challenge 32, 33. In our model, the immunodominant CD8+ T-cell epitope was found to be cross-reactive, but to differing degrees, following either JEV or WNV infection. Our detailed characterization of these epitope-specific responses did not demonstrate an alteration in epitope hierarchy, but rather differences in cytokine profiles and T-cell phenotype. As previous studies have elucidated a role for subdominant cross-reactive CD4+ and CD8+ T cells in protection as well as immunopathology, future experiments will address find more the role of the two cross-reactive CD4+ T-cell epitopes we identified and subdominant cross-reactive CD8+ T-cell epitopes along with the immunodominant cross-reactive CD8+ T-cell epitope in secondary heterologous JEV and WNV infections 10, 11. Here, we have shown that primary infections with JEV and WNV give rise to functionally and phenotypically distinct CD8+ T-cell responses. These

differences are due to the infecting virus (JEV versus WNV) rather than the stimulating variant (WNV S9 versus JEV S9) or viral pathogenicity. The JEV/WNV cross-reactive CD4+ and CD8+ T-cell epitopes we have identified will be useful tools to study the pathogenesis of sequential heterologous flavivirus infections. Flaviviruses continue to emerge into new geographic regions of the world, giving rise

to the possibility of new patterns of sequential infection with unknown outcomes (e.g. WNV into dengue- and yellow fever virus-endemic regions of South America). Altered CD8+ T-cell effector functions between flaviviruses may to lead to immunopathology or protection upon a secondary flavivirus infection. Additional experiments are needed to determine whether cross-reactivity Silibinin occurs between other members of the flavivirus family and its possible impact on disease outcome. JEV strain SA14-14-2 was provided by Dr. Thomas Monath (Acambis, Inc.). JEV strain Beijing was provided by Dr. Alan Barrett (University of Texas Medical Branch, Galveston, TX, USA). WNV strain 3356 was provided by Dr. Kristen Bernard (Wadsworth Center, Albany, NY, USA). Flaviviruses were propagated and titered in Vero cells (ATCC). The EL-4 T-cell lymphoma cell line (H-2b) served as target cells. Peptide (15–19mer) arrays corresponding to the entire proteome of WNV were obtained through the NIH Biodefense and Emerging Infections Research Resources Repository, NIAID, NIH (BEI Resources, Manassas, VA, USA). Peptide truncations (>70 or >90% purity) were obtained from AnaSpec (San Jose, CA, USA) and 21st Century Biochemicals (Marlborough, MA, USA).

Specifically, it was found that addition of RA during Flt3L-driven DC development skewed the culture drastically toward the SIRPαhi CD11bhi DC subtype, although it was not examined whether ESAM was also expressed under these conditions. Whether the concomitant reduction in the culture equivalents of the CD8hi/XCR1hi lineage (SIRPαlo CD11blo), and the reduced proportion of CD8+ DCs in mice kept on a vitamin A-rich diet indicates that RA also actively represses the development of CD8hi/XCR1hi DCs is an exciting possibility that remains to be tested. Taken together Beijer et

al. [13] demonstrate that RA signaling is necessary for the development of the ESAMhi CD11bhi DC subset within this website the spleen. These insights bring into focus a number of questions. For example, at what developmental stage are DCs subjected to RA conditioning? In this regard, it is interesting to note that hematopoietic stem cells in the bone marrow are among the few leukocytes that produce RA [19]. Analogous to the exposure of CD103+ DCs to RA in the intestine rather than in lymphoid organs [15], it is possible that pre-DCs would be exposed to RA in the bone marrow

prior to their migration Trametinib price toward the spleen (Fig. 1A). Given that the proportion of circulating pre-DCs was unaffected in the absence of vitamin A, such a conditioning effect of RA would have to be regulated at a qualitative level rather than simply by altering the precursor product relationship numerically. Nevertheless, whether these events take place at the pre-DC level in the bone marrow or occur in the spleen requires further investigation. Another consideration arising from these findings relates to the potential mechanism by which RA signaling promotes the development of ESAMhi CD11bhi DCs. Given that ESAMhi CD11bhi DC differentiation is dependent upon Notch 2 receptor signaling, an interesting explanation maybe linked to RA

regulating the expression of Notch 2 receptor and/or its ligands (Fig. 1B). PAK5 Indeed, it has recently been demonstrated that Raldh2-deficient mice, which are unable to convert vitamin A to RA, have diminished Notch signaling in developing neural tissues [20]. Thus, it is possible that RA signaling directly drives the expression of Notch 2 receptor and/or the ligands, and in this fashion, is necessary for the development of ESAMhi CD11bhi DCs. It will be interesting to examine whether the RA-dependent DC subsets express differential levels of Notch 2 receptor relative to other DC subsets. Similarly, future studies should investigate whether the Notch 2 receptor and/or its ligands contain RA-responsive elements within their promoter regions and are thereby directly regulated by RA. An alternative, although not mutually exclusive, mechanistic explanation may be that RA conditions pre-DCs to localize to distinct areas within the spleen that favor the differentiation of ESAMhi CD11bhi DCs (Fig. 1B).

The slides were Ruxolitinib then washed and incubated with TRICT-conjugated secondary antibody for 2 h. Anti-I-Ad

antibody was used after direct labelling with Alexa Fluor® 488 (Invitrogen, Carlsbad, CA, USA). Finally, the cells were counter-stained with DAPI. After a final wash, the slides were mounted in anti-fade solution [2.5% DABCO, 200 mm Tris–HCl (pH 8.6) and 90% glycerol], covered and sealed. Microscopic observation was performed using a confocal laser scanning microscopy (LSM 510 META, Carl Zeiss, Thornwood, NY USA). The full-length pro-IL-16 gene was initially obtained from 38B9 cells through a pro-IL-16-specific reverse transcriptase-polymerase chain reaction (RT-PCR). The product was eluted and then cloned into the pGEM®-T easy vector system (Promega). After enzyme digestion (BamHI/SalI), the cleaved gene was inserted into the pcDNA3.1 (+) mammalian expression vector (Invitrogen). Either control pcDNA3.1(+) or pro-IL-16/pcDNA3.1(+) DNA was mixed with 4 μl lipofectamine 2000 (Invitrogen) and incubated at room temperature for 20 min before being applied to the cells (5 × 106 cells/500 μl in a 24-well plate). At 24 h after transfection, the medium was changed and transfected cells were selected in G418-containing medium for 2 weeks. Three Stealth™ siRNA fragments for mouse pro-IL-16 (GenBank accession number:

BC026894; #1: 5′-CCU UGG IDH inhibitor cancer GUU AGA AUU UCC GAC UGC A-3′; #2: CAG GCA GAG AAU CAG CUC CUU UGA A-3′; #3: GAC CAG GUG UCA AGA UGC CAA GUC A-3′) and a Stealth™ RNAi negative Ketotifen control duplex (medium GC) were obtained from Invitrogen. Low-conductivity electroporation pulse medium (siPORT siRNA electroporation buffer) and GAPDH, as a positive control, were purchased from Ambion (Austin, TX, USA). To transfect the siRNA transiently, 38B9 (5 × 105) cells were centrifuged at 300× g for 6 min, and the cell pellets were resuspended in 75 μl pulse medium. Cells were then incubated with 1.5 μg siRNA and transferred into a 1-mm electroporation cuvette

(Bio-Rad) and immediately pulsed using a Gene Pulser® II electroporation system (Bio-Rad). Electroporation conditions were 120 mV, 500 μF and 100 Ω. After electroporation, the cells were incubated in a cuvette at 37 °C for 10 min and then transferred into prewarmed growth medium. The cells were used for subsequent analysis 40 h after transfection. To isolate total RNA, 38B9 cells (1 × 106) were harvested and washed in PBS, and total RNA was isolated using the easy-BLUE™ total RNA extraction kit (Intron Biotechnology, Sungnam, Korea). The purity and concentration of total RNA were measured using a SmartSpec™ Plus spectrophotometer (Bio-Rad). Five micrograms of RNA were reverse transcribed (Promega) to synthesize cDNA. For PCR amplification, each 25 μl reaction mixture contained 10 pmol each of forward and reverse primers, 5 μl cDNA and 0.25 U of Go Taq® DNA polymerase (Promega).

In addition, detailed assessment of the potential donor’s family history, presence of haematuria in family members, and extrarenal manifestations of Alport syndrome may help identify potential donors at risk of having underlying subclinical disease. There are no studies that have properly examined the issue of haematuria in live kidney donors. Most of our information Crenolanib supplier comes from studies of the incidence of haematuria in the general population and from the known pathological associations with this finding. Case reports exist in the literature, describing donors with known glomerular abnormalities with good short-term outcomes for donor and recipient. No large, prospective,

controlled studies have been performed. British Transplant Society / British Renal Association: An extensive, 100-page document has been produced outlining similar issues to those discussed here.

The full version of these British Live Donor Guidelines is available at: http://www.bts.org.uk and at http://www.renal.org Persistent microscopic haematuria in the potential living donor requires full investigation Gefitinib to identify an underlying cause, up to and including renal biopsy if there is no obvious urological explanation. Where there is insufficient evidence to quantify the risks following histological diagnoses of renal pathology, donation is not recommended. The Amsterdam Forum: A short manuscript outlining similar issues to those discussed here. Isolated microscopic hematuria (defined as 3–5 urinary sediment red blood cells (RBCs)/HPF) may not be a contraindication to donation. RBCs with glomerular origin have a dysmorphic appearance observed by phase-contrast microscopy and automated RBC analysis. Patients with persistent microscopic hematuria should not be considered for Sinomenine kidney donation unless urine cytology and a complete urologic work up

are performed. If urological malignancy and stone disease are excluded, a kidney biopsy may be indicated to rule out glomerular pathology such as IgA nephropathy. European Renal Association-European Dialysis and Transplant Association: (Nephrol Dial Transplant 2000): Exclusion criteria include: ‘reduced GFR (in comparison to normal range for age), proteinuria >300 mg/day, microhematuria (except when a urologic evaluation and possible kidney biopsy are normal), or hypertension without good control’. 1 Prospective, controlled studies on long-term living kidney donor outcomes, including an assessment of the utility of tests for haematuria and outcomes of donors with isolated urinary abnormalities such as microscopic haematuria. John Kanellis has no relevant financial affiliations that would cause a conflict of interest according to the conflict of interest statement set down by CARI.

Cells were cultured in RPMI-1640 supplemented with 10% heat-inactivated fetal calf serum, 100 U/ml penicillin,

100 mg/ml streptomycin, 50 μg/ml gentamicin and 2 mm l-glutamine (all from Invitrogen, Eugene, OR) at 37° in a humidified 5% CO2 incubator. Purified CD4+ subsets were activated in the presence of anti-CD3 antibody (purified OKT3 0·5 μg/ml) and autologous PBMCs irradiated with 40 Gy gamma-radiation, as a source of multiple co-stimulatory ligands provided BGB324 concentration by B cells, dendritic cells and macrophages found in these populations.28 In other experiments, cells were cultured in the presence of recombinant human (rh) IL-2 (5 ng/ml), Trichostatin A mouse IL-7 (10 ng/ml) or IL-15 (5 ng/ml) (all from R&D Systems, Minneapolis, MN). Cytokines were added at the beginning of the cell culture and not replenished. These cells were harvested at different times for phenotypic

and functional analyses. The PBMCs were stimulated with 10 μg/ml of purified protein derivative (PPD; Statens Serum Institut, Copenhagen, Denmark), 1/50 dilution of varicella zoster virus (VZV) -infected cell lysate, 1/200 dilution of Epstein–Barr virus (EBV) -infected cell lysate or 1/50 dilution of herpes simplex virus (HSV) -infected cell lysate (all from Virusys, Taneytown, MD). A CMV-infected cell lysate (used at 1/10 dilution) was prepared by infecting human embryonic lung fibroblasts with the Towne strain of CMV (European Collection of Animal Cell Cultures) at a multiplicity of infection

of 2. After 5 days, the infected cells were lysed by repeated freeze–thaw cycles. The PBMCs were left unstimulated or stimulated with antigenic lysates for 15 hr at 37° in a humidified CO2 atmosphere, with 5 μg/ml brefeldin A (Sigma-Aldrich) added after 2 hr. The cells were surface stained with peridinin chlorophyll protein-conjugated (-PerCP) CD4, phycoerythrin-conjugated PLEKHB2 (-PE) CD27 and phycoerythrin-Cy7-conjugated CD45RA (BD Biosciences) on ice. After being fixed and permeabilized (Fix & Perm Cell Permeabilization kit; Caltag Laboratories, Buckingham, UK), cells were stained with allophycocyanin-conjugated (-APC) interferon-γ (IFN-γ). Samples were acquired on an LSR I flow cytometer (BD Biosciences). For bone marrow experiments, paired peripheral blood and bone marrow samples were stimulated and analysed in parallel.

Furthermore, repeated sequences from the same individual can vary in copy number in different organs and tissues [16]. The general mechanisms

that lead to changes in copy number include homologous recombination and non-homologous repair mechanisms [17]. Changes in copy number might alter the expression levels of genes included in the CNVR. For example, the salivary amylase gene, AMY1, shows CNV in human populations, and the amount of salivary amylase is directly proportional to the copy number of AMY1[18]. More importantly, CNVs shape tissue transcriptomes on a global scale [19]. Additional copies of genes also provide redundancy that allows some copies to evolve new or modified functions while other copies maintain the original function. CNVs can represent benign polymorphic variations or convey clinical phenotypes by mechanisms such as altered gene dosage and gene disruption. CNV selleck products can be responsible for sporadic birth defects [20], other sporadic traits, Mendelian diseases and complex traits including autism, schizophrenia, epilepsy, Parkinson

disease, Alzheimer disease, human immunodeficiency virus (HIV) infection and mental retardation [21–23]. Interestingly, the set of genes that vary in copy number seems to be enriched for genes involved in olfaction, immunity and secreted proteins [24]. The following diseases are associated with CNVs of the immune genes: (i) CNVs of FCGR3B and FCGR2C (encoding different Fcγ receptors) have been associated with a range of autoimmune diseases, including selleck kinase inhibitor systemic lupus erythematosus (SLE), polyangiitis, Wegener’s granulomatosis and idiopathic thrombocytopenic purpura [25–27]. (ii) CNVs of the complement genes CFHR1 and CFHR3, which belong to the complement factor H protein family, have been associated with age-related macular degeneration and atypical haemolytic-uraemic syndrome [28–30]. Complement C4 gene copy number has been related directly

with systemic lupus erythematosus (SLE) [31]. (iii) On chromosome 8, a unit of seven β-defensin genes, which encode anti-microbial peptides with other diverse functions such as chemokine activity [32], has variability in its copy number [33]: low copy number has been associated with Crohn’s disease [34,35], and high copy number with predisposition to psoriasis [36]. (iv) In tuclazepam this review, we will examine one of the most striking examples of CNV in the human genome, the chemokine genes CCL3L and CCL4L. Chemokines are a large superfamily of small structurally related cytokines that regulate cell trafficking of various types of leucocytes to areas of injury, and play key roles in both inflammatory and homeostatic processes. Chemokines are classified into four families based on the arrangement of the first two cysteines of the typically conserved four cysteines: CXC, CC, C and CX3C (where X is any amino acid) [37].

Since total numbers of migrated CD4+ T cells did not differ between HD and RR-MS samples, lower Treg percentages under non-inflammatory this website conditions can be excluded to be due to increased migration of non-Treg. In line with our data on murine Treg transmigration, human HD Treg displayed consistent basolateral accumulation in the absence of endothelial cells. Higher Treg motility compared to non-Treg has previously been suggested as a mechanism of suppression of T effector cell function

as Treg were shown to be superior to TH cells in establishing close contact to dendritic cells, subsequently inhibiting their full maturation 27. Our finding of an augmented Treg motility in HD therefore is very well in line with this previous data. Furthermore, our observation of a migratory dysfunction of MS patient derived Treg introduces the idea that the presumed “regulatory deficiency” of CD4+ Treg in MS could at least be partially due to impairment in Treg motility. Our study provides first evidence of augmented overall cell motility as a constitutive feature of both R428 datasheet murine and human naturally occurring regulatory T cells. Adhesion ligand and chemokine receptor patterns expressed by Treg and their non-regulatory counterparts presumably determine

site-specific homing and have recently been a matter of substantial interest. Their innate cell motility, however, forms the basis of transendothelial diapedesis to and locomotion within any tissue and has been completely neglected in the past. Our data demonstrate Hydroxychloroquine an innate migratory superiority of murine and human Treg over naïve non-Treg. This migratory advantage should contribute to the role of Treg in maintaining tissue immune homeostasis and CNS immune surveillance.

However, this can be disturbed under conditions of autoimmunity, as demonstrated for MS patient-derived Treg. Albeit speculative, our findings could have relevance for the understanding of early lesion development and remitting phases during MS course. Twelve patients (9 female, 3 male) suffering from clinically definite RR-MS according to the revised McDonald diagnostic criteria 28 were enrolled in this study. All patients were in a stable phase of the disease, with relatively low scores on Kurzke’s expanded disability status scale (EDSS<3.5) and neither currently nor previously receiving any immunomodulatory treatment (age: 41.7±12.6 years, disease duration: 4.9±6.6 years, EDSS: 1.4±0.8). Ten HD (7 female, 3 male) with no previous history of neurologic disease served as controls (age: 34.1±12.2 years). There was no significant difference in age and gender distribution between patients with MS and healthy individuals. The study was approved by the local ethics committee and informed written consent was obtained from all participants. Six-wk-old female C57BL/6 mice were obtained from Harlan Laboratories.

2a). The B220+ CD43− fraction can be further subdivided based on surface IgM and IgD expression into pre-B (IgM− IgD−), immature

(IgM+ IgD−) or mature (IgM+ IgD+) B cells29 (Fig. 2a). We found that WT and dnRAG1 mice exhibited Deforolimus concentration a similar percentage and absolute number of B220+ CD43+ B cells, but the more mature B220+ CD43− B-cell subset was slightly lower in dnRAG1 mice compared with WT mice because of a significant reduction of mature B cells (Fig. 2a,b; see Supplementary material, Table S2). Taken together, these data suggest that dnRAG1 expression impairs B-cell development in the bone marrow at the immature-to-mature B-cell transition. Upon reaching the immature stage, B cells migrate to the spleen to complete their maturation, progressing through phenotypically and functionally distinct transitional stages during this process.30,31 Splenic B220hi B cells can be initially segregated based on the differential expression of AA4.1 (CD93) into transitional (B220hi AA4.1+) and mature (B220hi AA4.1−) subsets. Transitional cells can be further classified into subsets based on the 17-AAG manufacturer differential expression of surface IgM and CD23.32 T1 B cells (IgMhi CD23lo) are considered as immature B cells that have recently emigrated from

the bone marrow, which can differentiate into T2 B cells (IgMhi CD23hi).32 A third transitional B-cell subset, T3 (IgMlo CD23+), is thought to consist of immature B cells that have been rendered anergic by encounter with self-antigen.31,33 The mature B-cell population can be further subdivided by the differential expression of CD21 and CD23

into follicular (CD21int CD23−) and marginal zone (MZ; CD21hi CD23+) B-cell subsets.31 Consistent with observations in the bone marrow, dnRAG1 mice exhibit a significant reduction in the number of splenic transitional (B220hi AA4.1+) B cells compared with WT mice, because of a significant loss of cells in the T2 and T3 subsets (Fig. 2a,b; see Supplementary material, Table S2). In dnRAG1 mice, the mature B220hi AA4.1−subset is also significantly reduced relative to WT mice, with most of the difference attributed Flucloronide to a significant decrease in follicular B cells, but not MZ B cells (Fig. 2a,b). To explain the lack of an apparent defect in early B-cell maturation and in T-cell development in dnRAG1 mice, we used qPCR to detect total RAG1 transcript in various tissues and compare the relative abundance of RAG1 transcript between normal and dnRAG1 mice after normalizing to an internal calibrator (β-actin). From these experiments, we found that splenic RAG1 transcript levels are about 120-fold higher in dnRAG1 mice compared with normal littermates, but little difference was observed in thymus, bone marrow, lymph node, or liver (Fig. 3a,b).