Differences in the soluble HLA-G blood serum concentration levels in patients with ovarian cancer and ovarian and deep endometriosis. Am J Reprod Immunol 2010 Problem  The relationship between endometriosis and cancer has been widely discussed in the literature but is still not well clarified. Perhaps significantly, soluble human leukocyte antigen-G (sHLA-G) has been identified in the microenvironment of both ovarian cancer and endometrioma. The aim of this study has been to evaluate the sHLA-G levels in the blood sera of women with deep endometriosis and ovarian endometrioma

over the course of the menstrual cycle and to compare to the levels of sHLA-G in the blood sera of women with ovarian Midostaurin nmr cancer. Method of study  In our study, we examined the blood sera obtained from 123 patients operated on because of ovarian cancer (65 cases), ovarian endometrioma (30 cases), and deep endometriosis (28 cases). We decided to compare the levels of sHLA-G in Selleck EPZ6438 patients with endometriosis to those found in patients with ovarian cancer with respect to the menstrual cycle phases. The sHLA-G concentration level was measured by enzyme-linked immunosorbent assay kit. Results  The level of sHLA-G concentration in the blood serum of patients with deep endometriosis fluctuates over the course of the menstrual cycle, and during the proliferative and secretory phases,

it remains at a high level comparable to that found in patients with ovarian cancer. By contrast, the level of sHLA-G

concentration in the blood serum of patients with ovarian endometrioma fluctuates minimally over the course of the different menstrual cycle phases and, as in patients with ovarian cancer, it remains at high level during the proliferative phase. Conclusion  sHLA-G blood serum concentration levels would seem to provide important information regarding the degree of immune system regulation disturbance in both ectopic endometrial cells and the cancer cell suppressive microenvironment. “
“The role of mast cells (MCs) in Bay 11-7085 the generation of adaptive immune responses especially in the transplant immune responses is far from being resolved. It is reported that mast cells are essential intermediaries in regulatory T cell (Treg) transplant tolerance, but the mechanism has not been clarified. To investigate whether bone marrow-derived mast cells (BMMCs) can induce Tregs by expressing transforming growth factor beta 1 (TGF-β1) in vitro, bone marrow cells obtained from C57BL/6 (H-2b) mice were cultured with interleukin (IL)-3 (10 ng/ml) and stem cell factor (SCF) (10 ng/ml) for 4 weeks. The purity of BMMCs was measured by flow cytometry. The BMMCs were then co-cultured with C57BL/6 T cells at ratios of 1:2, 1:1 and 2:1. Anti-CD3, anti-CD28 and IL-2 were administered into the co-culture system with (experiment groups) or without (control groups) TGF-β1 neutralizing antibody.

Screening based on title and abstract identified 56 citations for full-text review (Fig. 1). Additional five studies[25-27, 39, 53] were identified from reference lists of the identified articles and from other databases. Of the 56 potentially relevant articles,

32 were excluded for reasons given in Figure 1, leaving a total of 24 studies[24-47] that met the inclusion criteria. Twenty one studies[24-30, 32, 34-38, 40-47] reported associations GS-1101 cell line between use of statins and AKI, and 14 studies[28, 31-35, 37, 39-41, 43-46] reported associations between use of statins and AKI requiring RRT. Five studies[24-28] used RCT design, and the rest applied a cohort design.[29-47] Only one RCT[28] defined AKI as the primary endpoint. The other four RCTs defined postoperative thrombocytosis,[24] postoperative inflammatory responses,[25]

postoperative myocardial injury,[26] and the number of postoperative endothelial progenitor cells[27] as primary endpoints. Among the cohort studies, only three used prospective design; the remaining studies were retrospective in design. As for the study population, two studies involved nation-wide populations, while most of the other studies were conducted at one single centre. Among the two population-based studies, one was conducted in Canada,[43] and the other in the USA.[47] We assessed the quality GSK-3 phosphorylation of included studies with the Jadad scale.[54] The study conducted by Prowle JR and colleagues[28]

had the highest score on the Jadad scale. The results were summarized in the Appendix 1 (Table App1). The studies varied in their types of surgery, mean age, and case definition (Table 1). The types of surgery were restricted to cardiac or vascular surgery in most studies. Specific type, dosage, and duration of preoperative statin therapy until were not available in most studies. In contrast to AKI defined by database codes, AKI defined by a pre-specified increase of serum creatinine level was regarded as ‘AKI defined by laboratory criteria’. Among these, there were seven studies[28, 37, 38, 41, 44-46] using AKIN or RIFLE criteria[48, 49] as the definition for AKI. In all studies, the definition of AKI requiring RRT was based on clinical judgment without additional objective laboratory criteria. Specific statin type available i Dosage and duration not available Increase of sCr level > 30% (AKIN stage 1) Atorvastatin 20 mg/day or simvastatin 20 mg/day for at least 6 months Started before surgery Type, dosage and duration not available At least one dose of statin between admission and surgery In the 21 studies reporting the association of statin use and AKI, the incidence of AKI ranged from 1.88%[43] to 52.17%[44] (Table 1). The pooled incidence of AKI for all 21 studies was 4.89%. The pooled incidence of AKI among statin user and nonstatin user were 6.13% and 4.28%, respectively (Table 2).

Endometrial biopsy, endocervical curettage, cytobrush, and blood were collected during mid-luteal phase from 23 healthy

women. T-cells were isolated and analyzed by flow cytometry. As compared with their counterparts in blood, endometrial and endocervical T-cells had enhanced CCR5 expression, and were enriched for activated, effector memory cells. Endometrial T-cells were more responsive to polyclonal stimuli, producing a broad range of cytokines and chemokines. These findings underscore the responsiveness of endometrial T-cells to stimulation, and reveal their activated phenotype. These findings also suggest susceptibility of the upper reproductive tract to HIV-1 infection. “
“The thymic SCH772984 cell line medulla provides a microenvironment where medullary thymic epithelial cells (mTECs) contribute to the establishment of self-tolerance by the deletion of self-reactive T

cells and the generation of regulatory T cells. The progression of thymocyte development critically regulates the optimum formation of the thymic medulla, as discussed in this article. Of note, it was recently identified that RANKL produced by positively selected thymocytes plays a major role in the thymocyte-mediated medulla formation. Indeed, transgenic expression of soluble RANKL increased the number of mTECs and enlarged the thymic medulla in mice. The effects of RANKL on the thymic medulla may be useful for the engineering of self-tolerance FER in T cells. Most T cells are generated in the Deforolimus order thymus and such a T-cell development is initiated within the microenvironment of the thymic cortex 1, 2. Immature thymocytes are induced by DLL4 and IL-7 to express the TCR, as well as the co-receptors CD4 and CD8 3, 4. A virgin repertoire of TCRαβ-expressing CD4+CD8+(DP) thymocytes is selected for an immunocompetent, i.e. self-protective and useful, repertoire in the thymic cortex. The positive selection of the

immunocompetent repertoire seems to rely on the repertoire of self-peptides that are uniquely expressed by cortical thymic epithelial cells (cTECs) 5, 6. Positive-selection-inducing TCR signals in DP thymocytes not only support the survival and differentiation of DP thymocytes into CD4+CD8− or CD4−CD8+ (single-positive, SP) thymocytes, but also activate cellular machineries that further promote repertoire selection in the thymic medulla. These machineries include an increase in the expression of chemokine receptor CCR7 on positively selected thymocytes. Given that the CCR7 ligand chemokines, CCL21 and CCL19, are strongly expressed in the thymus by medullary thymic epithelial cells (mTECs), the CCR7-expressing positively selected thymocytes are attracted from the cortex to the medulla 7–9. The medullary microenvironment of the thymus plays an essential role in the establishment of self-tolerance.

Co-administration of S. enterica serovar Typhimurium expressing swIL-18 and swIFN-α produced enhanced Th1-biased humoral and cellular immune responses against see more the inactivated PrV vaccine, when compared to single

administration of S. enterica serovar Typhimurium expressing either swIL-18 or swIFN-α. Also, enhanced immune responses in co-administered piglets occurred rapidly after virulent PrV challenge, and piglets that received co-administration of S. enterica serovar Typhimurium expressing swIL-18 and swIFN-α displayed a greater alleviation of clinical severity following the virulent PrV challenge, as determined by clinical scores and cumulative daily weight gain. Furthermore, this enhancement was confirmed by reduced nasal shedding of PrV following viral challenge. Therefore, these results suggest Nutlin-3a manufacturer that oral co-administration of S. enterica serovar Typhimurium expressing swIL-18 and swIFN-α provide enhanced Th1-biased immunity against inactivated PrV vaccine to alleviate clinical signs caused by PrV challenge. The

combined administration of two or more cytokines may produce effects that are antagonistic, additive, or synergistic (1). The potential effectiveness of cytokine combinations has been addressed empirically, based upon mechanisms to determine the nature of innate and acquired immunity (2–4). Among such effects, additive and synergistic effects may be useful when immunizing hosts. The enhanced Sulfite dehydrogenase effects of cytokine

combinations for immunomodulation or antiviral activity have been evaluated in several infectious diseases of livestock animals such as FMD (5,6), PRRS (7), and PrV (8). The outstanding feature of interferon-alpha (IFN-α), which is a type I IFN, is its ability to nonspecifically inhibit viral growth by inducing the expression of numerous cellular genes through interaction with specific type I receptor complexes and triggering of the Janus-activated kinases (JAKs)-signal transducer and activators of transcription (STAT) 1/2 pathways (9). Interleukin 18 (IL-18), originally known as IGIF, can act on T helper 1 (Th1) cells, non-polarized T cells, NK cells, B cells, and DCs to produce IFN-γ in the presence of IL-12, through specific IL-18R complexes and triggering of MyD88-IRAK-TRAF (10). In addition, virus-infected macrophage-derived IL-18 and type I IFN (IFN-α/β) produced by the same cells synergistically induce rapid IFN-γ production, leading to the induction of Th1 immune responses (11). Therefore, type II IFN-γ induced by IL-18 may act synergistically with type I IFN to produce enhanced modulation of immune responses against specific antigens (5). Pseudorabies virus (PrV) is an alpha-herpes virus that causes a fatal illness in swine known as Aujeszky’s disease.

Remove supernatant completely. Critical troubleshooting! This step is the primary cause of non-specific positive results with the secretion assay. Centrifuging cells into a pellet when they are still

warm will contaminate the assay. Keep the cells ice-cold to stop secretion AZD8055 price of cytokines after the secretion period. Ensure that the wash is in buffer, as the ethylenediamine tetraacetic acid (EDTA) helps to stop the reaction Repeat washing step in ice-cold buffer. During the second wash prepare the cytokine detection antibody. This is diluted by adding 20 µl of cytokine detection antibody stock to 80 µl of ice-cold buffer; 100 µl of this stock solution is required per 1 × 107 cells. For example, for 5 × 107 dilute 100 µl of reagent with 400 µl buffer. Store on ice until used. For detection of two cytokines, add 10 µl of each detection antibody per 80 µl buffer.

Critical step– if separating two cytokine populations consecutively, add only one anti-fluorochrome microbead at this point. The second microbead can be added after the first separation: repeat the steps described here. Completely remove supernate. Resuspend in 500 µl buffer. For magnetic labelling, add 100 µl diluted anti-PE or APC microbeads per 1 × 107 cells, mix well and incubate for 15 min at 8°C (i.e. in the refrigerator, not on ice). Critical step– it is essential to have an unseparated sample to work out the start frequency and subsequent recovery of cells. Prepare two MS columns per sample by rinsing I-BET-762 ic50 with 500 µl of cold buffer. Place the first column into the magnetic field of a suitable MACS Separator (e.g. MiniMACS). Troubleshooting – it is essential to use two columns. Each column

can enrich the cells about 100 times. Thus, because of the low frequency of cytokine-producing cells, two columns are required to obtain the best unless purity. If cells are to be cultured: if cells are to be cultured directly after isolation, cells can also be eluted with culture medium. In this case, replace the last buffer wash with a medium wash, and then elute the cells with medium. If medium is to be used, ensure that it does not contain any particles, e.g. from serum. If in doubt, filter medium before use. If medium elution is used, cells for flow cytometry should be washed free of any phenol red. Critical point.  Do not use any PE- or APC-based tandem fluorochromes to stain cells sorted with anti-PE or APC beads, as they will be bound and stain non-specifically. All cytokine assays are low-frequency analyses. To properly identify cytokine producing cells, both positive staining with, e.g. CD4 or CD8 is required and also exclusion of unwanted cells from the analysis is vital. Exclusion of the dead cells (lymphocyte gating alone is not enough) using propridium iodide (PI), 7-AAD or other vital dyes will virtually eliminate non-specific background staining. It may also be necessary to exclude cells that tend to non-specifically bind fluorochromes, e.g.

This was achieved by stirring one volume of 2% (w/v) alginate solution for 20 min with one-half PS-341 price volume of 0·08% (w/v) 1-Ethyl-3-(3-dimethyllaminopropyl)carbodiimide hydrochloride (EDC-HCl) and 3% (w/v) sulfo-NHS solution. The resulting mixture was incubated for 17 h at room temperature with one volume of

alfa-t-butyloxycarbonylamino-omega-amino poly (ethylene glycol) PEG (MW 5000 Da). After dialysis using a tubular membrane (100 kDa MWCO, Spectra/Por® Biotech Cellulose Ester; Spectrum Ls Europe B.V., Breda, The Netherlands) against 1000 volumes of demineralized water, the product was freeze-dried, weighed and placed in flat bottom beaker to be completely covered for 40 min at room temperature by trifluoroacetic acid (TFA; Fluka Sigma-Aldrich Ltd). Thereafter, the TFA was removed under a nitrogen

flow, and the product finally freeze-dried overnight. The alginate-PEG5k-NH2 (modification rate 1 : 50 units) obtained was dissolved at 2 mg/mL in carbonate buffer 0·1 m pH 9·0 (freshly prepared). One volume of 0·1% (w/v) α-d-mannopyranosyl-phenyl isothiocyanate (Fluka Sigma-Aldrich Ltd) in DMSO was then added drop-wise with constant agitation to 50 volumes of alginate (theoretical modification rate was 1 : 50 units). After approximately 30- min agitation, the solution was stored overnight at 4°C. The suspension was then dialysed with a 100 kDa MWCO membrane (Spectrum Ls Europe B.V.) against 300 volumes demineralized H2O. The filtrate was changed four times every 2 h, and the product freeze-dried Crizotinib ic50 for storing at −20°C. Mannose-alginate decorated nanogels were prepared as described in Nanogel surface decoration with alginate, using this alginate-mannose instead of alginate. The final concentration of recNcPDI in the nanogel suspension after concentration was 50 μg PDI/mL dispersion. Recombinant NcPDI and recNcPDI-nanogel preparations were subjected

to ultracentrifugation (150 000 × g, 25 min, 4°C) using a TST55.5 rotor and a Centrikom T-2070 ultracentrifuge. The association of recNcPDI antigen with the nanogels was evaluated by analysing supernatant and pellet fractions by 12·5% (w/v) sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE), carried out under reducing conditions following boiling of the Adenosine triphosphate samples in sample buffer (40). Protein bands were visualized by silver staining and Western blotting as previously described (40). For immunoblotting, rat anti-recNcPDI (18) diluted 1 : 1000 in PBS containing 0·3% (w/v) BSA was used. The secondary antibody was an anti-rat IgG alkaline phosphatase conjugate (Promega, Madison, USA), which was applied according to the instructions provided by the manufacturer. One hundred and thirty female Balb/c mice (6 weeks of age) were purchased from Charles River Laboratories (Sulzheim, Germany) and were housed under conventional day/night conditions according to the standards set up by the animal welfare legislation of the Swiss Veterinary Office.

Rather, the reported autoimmune deviations of cDC-less animals 13, 14 are related to their development of a chronic myeloproliferative disorder. Here, we established that expression of the costimulatory molecules CD80 and CD86 by cDC is required for peripheral Treg maintenance. As such, our studies complement a recent study demonstrating that cDC control Treg homeostasis in dependence of MHC II expression 13. Using

CD80/CD86 mutant animals and a strategy that restricts the B7 deficiency to cDC, we show here that cDC also have to provide a critical costimulatory signal to the Treg. Animals that constitutively lack cDC display features of systemic lymphocyte activation including hypergammaglobulinemia, the accumulation of CD62LloCD44hi T cells and an increased prevalence of Th17 and Th1 cells 14, 15. Ohnmacht et al. interpreted these findings as an indication find more of a general tolerance failure in these animals resulting in fatal autoimmunity 14. Furthermore, after establishing that cDC are required for Treg homeostasis, Darrasse-Jeze et al. suggested that the elevation MLN0128 purchase in Th1 and Th17 in cDC-depleted animals is a result of their impaired Treg compartment 13. However, as we recently reported 15, constitutive and conditional ablation of cDC triggers

a systemic elevation of the growth factor Flt3L causing a progressive nonmalignant myeloproliferative disorder. Here, we show that the feedback loop that links the peripheral cDC compartment to myelogenesis is not mediated through CD80/86 interaction since animals that exclusively harbored B7-deficient cDC

did not develop the myeloproliferation. We had previously interpreted the lymphocyte activation in cDC-depleted mice as a consequence of the systemic pathological accumulation of myeloid cells, rather than as a result of a breakage of adaptive immune tolerance. In support of this notion, we had despite major efforts failed to detect T-cell autoreactivity in these animals 15. Taking advantage of mice that harbor the cDC-restricted B7 deficiency and display a reduction of Treg without Farnesyltransferase associated myeloproliferation, we show in thid study that the Treg reduction resulting from impaired cDC/T-cell crosstalk does as such not result in lymphocyte hyperactivation. Rather than reflecting a tolerance failure or autoimmunity, our results suggest that the latter is a secondary consequence of the Flt3L-driven myeloproliferative disorder observed in cDC-deficient animals. This notion is supported by the fact that other animals displaying myeloproliferative disorders, such as IRF8-deficient mice, have also been reported to suffer from hypergammaglobulemias 23.

In addition, in the normally formed glomeruli there was a significant increase in size, indicative of glomerular hypertrophy and thus hyperfiltration. The variability in the proportion of abnormal glomeruli in the outer renal cortex between preterm infants suggests that there may be differences in haemodynamics, and/or other factors in the postnatal environment of the infant (such as

exposure to NVP-LDE225 ic50 nephrotoxic drugs, oxygen supplementation, mechanical ventilation and co-morbidities) which may be negatively impacting glomerulogenesis[3] (Fig. 1). In this regard, there is a major haemodynamic transition at the time of birth when blood pressure and heart rate are markedly elevated[10] and blood flow to the kidneys is increased.[11] Hence, it is possible that the developing capillaries of immature glomeruli are not prepared for the haemodynamic transition at birth and their formation is adversely affected. Indeed, we have recently shown that there is injury to the wall of the aorta as a result of preterm delivery.[12] see more In future studies, it is imperative to determine the cause of the glomerular abnormalities in the preterm kidney, in order to maximize the number of functional nephrons at the beginning of life; this will likely lead to short-term and long-term benefits to renal health. “
“We recommend that all candidates

for kidney transplant are screened for cardiovascular risk factors (1B). Indicators of high risk include (1B): Older age. Diabetes mellitus. Abnormal echocardiogram (ECG). Previous ischaemic heart disease or congestive heart

failure. Increased duration of dialysis. Smoker. We suggest that kidney transplant candidates with a low clinical risk of cardiovascular disease do not require stress testing for coronary artery disease (2B). We suggest that kidney transplant candidates with a moderate or high clinical risk of C1GALT1 cardiovascular disease undergo cardiac stress testing prior to transplantation (2B). The following should be noted in relation to cardiac stress testing in dialysis patients: Exercise ECG has a poor predictive value in patients on dialysis (2B). The use of a cardiac stress test such as dipyridamole thallium testing or stress echocardiography is predictive of significant coronary artery disease and major cardiac events in patients with higher clinical risk. Where possible we recommend that this testing should be performed without concurrent β-blocker therapy (1B). As the prognostic accuracy of cardiac stress testing in dialysis patients is of limited duration, it is suggested that testing be repeated in high risk patients. The interval at which testing should take place has not been well defined; however, the predictive value of a positive test diminishes after 24 months (2C). We recommend that coronary angiography be considered for kidney transplant candidates with abnormalities on screening procedures (1B). We suggest that the benefit of revascularization prior to transplantation be reviewed on an individual basis (2C).

Macaque and human pDC were shown to have similar TLR expression profiles [25], which is in agreement with the response patterns observed by us. Also TLR-7, TLR-9 and myeloid differentiation primary response gene 88 (MYD88) GS1101 sequences were shown to be identical, whereas there were important differences for interferon regulatory factor 7 (IRF-7) [26]. Other regulatory pathways still need to be explored [37]. Beside TLRs, the C-type lectin receptor (CLR) family plays an important role in the modulation of innate immune responses [38, 39]. Human pDC express the CLRs blood dendritic cell antigen 2 (BDCA2) and dendritic cell immunoreceptor (DCIR) [40]. Cross-linking of DCIR was shown to result in reduced IFN-α induction upon

TLR-9 stimulation [40], and similar inhibitory effects were reported following incubation with the CLR ligand mannan [41]. Interestingly, BDCA2 [our unpublished observation and documented at the NIH non-human primate reagent resource portal (http://nhpreagents.bidmc.harvard.edu/NHP)] and DCIR [42] were shown to be absent on pDC in rhesus macaques. Although not investigated here, a difference in the balance between activating TLRs and inhibitory CLRs could lead to different levels of pDC activation, possibly translating into a difference in cytokine production pattern. A direct comparison between the absolute numbers of pDC, mDC and monocytes in rhesus versus human blood showed that rhesus

macaques had a lower number of pDC, while Ensartinib research buy there was no difference in the abundance of the other subsets. The number

of pDC observed, i.e. 3020 ± 1357 cells/μl, is in agreement with several reports on rhesus macaques [16, 18, 24, 25, 43] and considerably less Amobarbital than in humans [44]. In contrast, two other studies, where a direct head-to-head comparison was made, showed no difference in pDC number [17, 28], although it must be noted that in those studies the quantification was either performed on PBMC or cynomolgus monkeys imported from Mauritius were used, which have a more limited genetic diversity and might differ from rhesus macaques. The strong IL-12p40 expression in rhesus pDC may have implications for preclinical evaluation of vaccines in this model. For instance, TLR-7/8 containing adjuvants might trigger different responses in macaques than in humans and involve pDC as IL-12 producing cells. Also TLR-9 agonists could be expected to induce an IL-12 response in rhesus macaques, in contrast to humans. Simultaneous production of IFN-α and the inflammatory cytokines TNF-α and T helper type 1 (Th1)-skewing cytokine IL-12 might also lead to a slightly different response pattern to bacterial and viral infection and have consequences for the induction of CD8 responses [45, 46]. We would like to thank Dr F. Verreck for critical reading of the manuscript, Dr S.B. Geutskens for organizing the collection of the human blood samples and H. van Westbroek for preparing the figures.

6) and when mouse CD11b+ spleen cells were used as effector cells (data not shown). When tested in different donors, the % shaving observed with mouse AT80 was typically between 20 and 47%, whereas other mouse antibodies induced shaving at 60–90%. We then tested related human or chimeric antibodies BHH2, CD20-2, CD20-6, CD20-G and chimeric AT80 (chAT80). However, here Small Molecule Compound Library we observed 67–84% shaving, which was comparable to the level observed with RTX (Fig. 7). Recently, it was reported that monocytes have an inhibitory effect on ADCC because they

can remove antibody such as RTX from the surface of target B cells and in this way cause a reduced ability of NK cells to bind RTX via the FcγRIII.11,12 Hence, monocytes seem to compromise RTX treatment, in particular in haematological malignancies with a large B-cell load.13 Here, we confirm these observations and demonstrate that the shaving mechanism is independent of endocytosis but relies on protease activity after monocyte binding to the Fc part of RTX. Also, we have screened a series of alternative type I and II anti-CD20 antibodies to identify antibodies with a reduced effect on monocyte-mediated shaving. This work demonstrated that monocytes are able to remove B-cell-bound RTX at monocyte : B-cell ratios of 1 : 2 in vitro and that this is dependent on the Fc part of RTX. Recent work has shown that the high-affinity receptor for IgG, FcγRI,

is responsible for this and expression of this receptor on monocytes provides a competitive advantage to hinder NK-cell-mediated ADCC through FcγRIII with lower affinity.12 This group also demonstrated that addition of human IgG could restore NK-cell-mediated ADCC Roxadustat in these co-cultures. However, in our assay, the addition of human IgG or anti-CD64 only had a minor effect on monocyte-mediated shaving. This could reflect that the addition of IgG in their assay had a direct effect on the NK cells, which also have an ability to perform shaving of target cells. Hence, monocytes could either be dependent on cross-linking

of even low numbers of free FcγRI to induce shaving or be activated in alternative ways. Interestingly, we also observed that Methisazone monocyte-derived dendritic cells can mediate the shaving reaction, and this could represent an additional mechanism whereby dendritic cells in the tumour microenvironment act as a ‘black hole’, hindering effective anti-tumour immune responses. Hyperosmolar sucrosis is an inhibitor of endocytosis. In our assay, hyperosmolar sucrosis did not lead to inhibition of the shaving reaction and this indicates that this phenomenon is not the result of B-cell-mediated endocytosis of the CD20/RTX complex or of simple endocytosis by monocytes. This observation is in line with detailed analysis from Beum et al.11 who recently demonstrated that the shaving reaction is similar to a processing mechanism originally described by Griffin et al.,10 which is now named trogocytosis.