[French Expert advice on the management of valproate in childbearing and pregnant women with bipolar disorder]. / Avis d'experts français sur la prise en charge des femmes en âge de procréer et enceintes souffrant d'un trouble bipolaire traitées par valproate.

INTRODUCTION: The perinatal period is associated with high risk of relapses in women with untreated bipolar disorder (BD) and can have significant consequences on foetal and child development. Valproate is an effective mood stabilizer in BD but it is also the anticonvulsant associated to the highest risks of neurodevelopmental disorders and congenital malformations. The National Agency for the Safety of Medicines and Health Products (ANSM) changed the conditions of use and prescription of valproate in France in 2015. Its prescription is now contraindicated (i.e., not to be prescribed) in women able to have children unless alternative treatments are ineffective or not tolerated. Moreover, valproate could only be prescribed if the protocol of a specific pregnancy prevention program is followed. METHODS: A panel of experts from the French Association for Biological Psychiatry and Neuropsychopharmacology (AFPBN) provided consensus-based recommendations for switching and discontinuation of valproate in women with BD. The development of these recommendations consisted of an adaptation to French clinical practice based on a European expert opinion published in 2019. The experts discussed five real-world clinical situations in light of the scientific evidence and their clinical experience (a. Stable BD patient with valproate monotherapy who is planning pregnancy, b. Stable BD patient with valproate polytherapy who is planning pregnancy, c. Unstable BD patient with frequent relapses and valproate polytherapy who is planning pregnancy, d. Stable BD patient treated with valproate and unexpected pregnancy, e. Unstable BD patient treated with valproate and unexpected pregnancy) and developed, through several rounds of exchange drafts, a French version of clinical recommendations. RESULTS: First of all, some factors need to be considered for establishing personalized practical recommendations for a safe and effective switching or discontinuation of valproate in any clinical situations: planned pregnancy or unplanned pregnancy or current pregnancy, the existence or not of a pregnancy risk minimization program and a complete treatment history. Other factors that should be considered are the predominant polarity, the severity, the stability, the comorbidities associated with BD, the beliefs toward treatments, the family situation and the preference of the patient. The modalities for switching or discontinuation of valproate in women with BD were related to the clinical situation. First-line therapeutic alternatives such as lithium, lamotrigine, quetiapine, olanzapine or aripiprazole were preferred for patients suffering from a clinically stable BD considering pregnancy or pregnant. In patients suffering from clinically unstable BD, to reach stability was considered first. A shared decision-making should be systematically implemented and the patient must be fully informed of the risks related to an in-utero exposure to valproate, and the risks of the discontinuation/switch that is considered. CONCLUSION: Although the adaptation to French practice of the recommendations from the European expert opinion highlighted some differences in the criteria taken into consideration to guide the therapeutic decision, this expert advice will guide the clinician for switching and discontinuation of valproate in BD women able to have children or pregnant.

[Erythrapheresis in children with sickle cell disease, experience of a paediatric apheresis unit]. / Érythraphérèses chez l'enfant drépanocytaire, expérience d'une unité d'aphérèse pédiatrique.

Erythrocyte exchanges on cell separators can be used in children with sickle cell disease and are effective in lowering the level of haemoglobin S. Of the 938 aphereses performed in 2020 in our unit, we observed a low rate of failure of procedures and few complications. Ninety-six percent of erythraphereses were performed in the context of chronic exchange programs, in more than 80% of cases for cerebral vasculopathy or after the occurrence of ischemic strokes. Less than 4% of the procedures were performed for specific indications (preparation for cholecystectomy most often). The vascular access is rarely an obstacle to the realisation of the apheresis. In case of insufficient venous capital, installing an arteriovenous fistula may be considered. Depending on the child's weight, haemoglobin level, and the severity of the sickle cell anaemia, precautions may be necessary when priming the procedure. Nurses experienced in paediatric apheresis and a good medical knowledge of sickle cell disease allowed us to use this technique from the age of 3years and the weight of 15kg.

Neuropsychology of traumatic brain injury: An expert overview.

Traumatic brain injury (TBI) is a serious healthcare problem, and this report is a selective review of recent findings on the epidemiology, pathophysiology and neuropsychological impairments following TBI. Patients who survive moderate-to-severe TBI frequently suffer from a wide range of cognitive deficits and behavioral changes due to diffuse axonal injury. These deficits include slowed information-processing and impaired long-term memory, attention, working memory, executive function, social cognition and self-awareness. Mental fatigue is frequently also associated and can exacerbate the consequences of neuropsychological deficits. Personality and behavioral changes can include combinations of impulsivity and apathy. Even mild TBI raises specific problems: while most patients recover within a few weeks or months, a minority of patients may suffer from long-lasting symptoms (post-concussion syndrome). The pathophysiology of such persistent problems remains a subject of debate, but seems to be due to both injury-related and non-injury-related factors.

Reproducibility of digital measurements of lower-limb deformity on plain radiographs and agreement with CT measurements.

INTRODUCTION: Five angles (HKA, HKS, alpha, beta, tibial slope) are used for goniometry in total knee arthroplasty. The reproducibility of the measurement of these angles has been assessed on plain and digitized x-rays, but to our knowledge, this has not been confirmed on x-rays taken on the PACS system and they have not been compared to computed tomography (CT) measurements, the reference for angle measurement. This prospective study aimed to: (1) evaluate the inter- and intrarater reliability of the measurement of these angles on digital x-rays taken on a PACS; (2) determine the agreement of these measurements with those obtained using a CT protocol. HYPOTHESIS: The measurements of these five angles on digitized radiographs are reproducible and in agreement with CT values. MATERIAL AND METHODS: Forty-two patients suffering from knee osteoarthritis and scheduled for total knee arthroplasty were included in the study. Each patient had a PACS digitized x-ray and a CT intended to produce patient-specific instrumentation (Symbios, Yverdon, Switzerland) including measurements of the angles evaluated. Four senior orthopaedic surgeon-raters measured all the angles twice. Inter- and intrarater reliability was then calculated as well as the agreement between the second measurement of each rater and the CT measurement using interclass correlation and kappa coefficients (data provided as means and 95% confidence intervals). RESULTS: The inter- and intrarater reliability values were excellent for the HKA, alpha, and beta angles (with, respectively, a coefficient of 0.99 [0.97-0.99], 0.84 [0.76-0.9], and 0.94 [0.86-0.96] interrater reliability and 0.98 [0.96-0.99], 0.86 [0.75-0.92], and 0.65 [0.44-0.8] intrarater reliability). Interrater reliability was low for HKS and tibial slope angles (coefficients all<0.4 for interrater reliability and <0.7 for intrarater reliability). The x-ray/CT agreement was very good for the HKA, alpha, and beta angles (0.81 [0.67-0.99], 0.74 [0.56-0.91], and 0.74 [0.45-0.92], respectively) and low for the HKS and tibial slope angles (all<0.45). DISCUSSION/CONCLUSION: The HKA, alpha, and beta angles were reproducible for digital radiographs and showed good agreement with CT measurements. HKS and tibial slope angles should be used with greater caution, and other navigation methods or patient-specific instrumentation should be explored. LEVEL OF EVIDENCE: Level III, prospective, comparative diagnostic case-control study.

Neurobehavioral and self-awareness changes after traumatic brain injury: Towards new multidimensional approaches.

Neurobehavioral and self-awareness changes are frequently observed following traumatic brain injury (TBI). These disturbances have been related to negative consequences on functional outcomes, caregiver distress and social reintegration, representing therefore a challenge for clinical research. Some studies have recently been conducted to specifically explore apathetic and impulsive manifestations, as well as self-awareness impairments in patients with TBI. These findings underlined the heterogeneity of clinical manifestations for each behavioral disturbance and the diversity of psychological processes involved. In this context, new multidimensional approaches taking into account the various processes at play have been proposed to better understand and apprehend the complexity and dynamic nature of these problematic behaviors. In addition, the involvement of social and environmental factors as well as premorbid personality traits have increasingly been addressed. These new multidimensional frameworks have the potential to ensure targeted and effective rehabilitation by allowing a better identification and therefore consideration of the various mechanisms involved in the onset of problematic behaviors. In this context, the main objective of this position paper was to demonstrate the interest of multidimensional approaches in the understanding and rehabilitation of problematic behaviors in patients with TBI.

Excellent short-term results of hip resurfacing in a selected population of young patients.

BACKGROUND: Hip resurfacing (HR) is an alternative option to total hip arthroplasty (THA) in a population of selected patients (young and/or active). HYPOTHESIS: The short-term survivorship rate is as least as good as that for THA with no abnormal increase in serum metal ion levels. MATERIALS AND METHODS: A continuous prospective series of 502 hip resurfacings in 481 patients mean age 48.7 years old (±10.3; 18-68) (Conserve Plus, Wright Medical Technology) was analyzed clinically, radiologically and biologically (total blood chrome, cobalt and titanium metal ion levels). Mean follow up was 4.1 years (1.9-4.9). RESULTS: There were no dislocations. There were 5 cases of revision surgery with component replacement (including 2 infections). Implant survivorship using implant removal as the criteria (excluding infection) was 99.4% at 4 years (CI 95%: 98.1-99.8). The evaluation of metal ion levels showed a significant increase in cobalt from a preoperative level of 0.24 µg/L (0.01-3.6) to 0.86 µg/L (0.01-5.7) at the final follow-up (P<0.001). Chrome and titanium levels went from 0.68 µg/L (0.01-4.4) and 2.36 µg/L (0.39-7) to 1.28 µg/L (0.1-5.5) and 4.49 µg/L (1.29-8.21) respectively (P<0.001). All clinical scores had significantly improved at the final follow-up. Mean frontal plane cup inclination was 42.7° (35-62). DISCUSSION: In a selected population of young and/or active patients, the short-term results of hip resurfacing are excellent. At the postoperative 4-year follow-up the rate of complications (in particular the absence of dislocations) was less than that for THA in young and/or active patients. Certain conditions must be respected to obtain these results; frontal plane cup inclination of between 40 and 45°, a femoral head diameter of at least 48 mm and good quality femoral bone. LEVEL OF EVIDENCE: IV.

Does severity of femoral trochlear dysplasia affect outcome in patellofemoral instability treated by medial patellofemoral ligament reconstruction and anterior tibial tuberosity transfer?

INTRODUCTION: Medial patellofemoral ligament (MPFL) reconstruction associated to anterior tibial tuberosity transfer (ATTT) is recommended in objective patellofemoral instability (PFI). Efficacy, however, has not been precisely determined in trochlear dysplasia with spur. A case-control study was performed in a PFI population, comparing groups with trochlear dysplasia with and without spur (S+ vs. S-) to assess the impact of trochlear dysplasia on (1) patellofemoral stability, (2) functional results and complications, and (3) patellofemoral cartilage status on MRI. HYPOTHESIS: Trochlear spur does not affect outcome in PFI managed by MPFL reconstruction and ATTT. MATERIAL AND METHODS: Twenty-eight knees (26 patients) with PFI were analyzed retrospectively and divided into 2 groups of 14 knees each according to presence of trochlear spur (S+ vs. S-). All 28 knees had undergone ATTT and MPFL reconstruction by semitendinosus autograft. Results were assessed on Lille and IKDC functional scores, and cartilage status was determined on MRI at last follow-up. RESULTS: At a mean 24 months' follow-up (range, 12-52 months), there was no recurrence of dislocation. IKDC and Lille scores tended to improve in both groups, although the only significant improvement was in IKDC score (S- gain, 21.3±16; S+ gain, 18.1±14) (P=0.01). IKDC scores at last follow-up were better in the S+ than S- group (79±19 [range, 21-92] vs. 68±13 [range, 35-84], respectively; P=0.012). Lille scores showed no significant inter-group differences in mean gain (P=0.492) or mean value (P=0.381). The S+ group showed more cartilage lesions (n=14/14 knees, including 12/14 with grade≥2 lesions) than the S- group (n=9/14 knees, all grade≤2). CONCLUSION: MPFL reconstruction with ATTT provided good short-term patellofemoral stability independently of the severity of trochlear dysplasia. Functional results and gain on IKDC, however, were poorer in case of dysplasia with trochlear spur. This is probably due to cartilage lesions, observed more frequently pre- and post-operatively in the spur group, especially as there was no significant difference in Lille Score, which highlights stability. LEVEL OF EVIDENCE: III, retrospective case-control study.

Computed tomography evaluation of hip geometry restoration after total hip resurfacing.

BACKGROUND: Anatomic reconstruction of the hip is among the main requirements for hip arthroplasty to be successful. Resurfacing arthroplasty may improve replication of the native joint geometry but has been evaluated only using standard radiographs. We therefore performed a computed tomography (CT) study to assess restoration of hip geometry after total hip resurfacing (HR), comparatively with the non-operated side. HYPOTHESIS: HR does not change native extra-medullary hip geometry by more than 5mm and/or 5°. PATIENTS AND METHODS: CT was used to evaluate unilateral HR in 75 patients with a mean age of 52.2years (range, 22-67years). The normal non-operated side served as the control in each patient. Mean follow-up was 2.5years (range, 1.9-3.1years). The primary evaluation criteria were femoral offset (FO) and femoral neck anteversion (FNA) and the secondary criteria were cup inclination angle, cup anteversion angle, and lower-limb length. RESULTS: FO showed a non-significant decrease (mean, -2.2mm; range, -4.5 to +3.7mm). FNA was preserved, with a difference of less than 2° at last follow-up versus the preoperative value. Cup measurements showed a mean anteversion angle of 24.8° (0.9-48.6) and mean inclination angle of 44.1° (32.1-56.3); corresponding values for the native acetabulum were 38.9° (20.5-54.8) and 24.8° (4.8-33.6). The residual lower-limb length discrepancy was less than 1mm (mean, -0.04mm [-1.2 to +1.6mm]). The mean angle between the femoral implant and the femoral neck axis was 5.4° of valgus. DISCUSSION: Our results show that HR accurately restored the native extra-medullary hip geometry. LEVEL OF EVIDENCE: III, prospective diagnostic case-control study.

Tantalum cones and bone defects in revision total knee arthroplasty.

Management of bone loss is a major challenge in revision total knee arthroplasty (TKA). The development of preformed porous tantalum cones offers new possibilities, because they seem to have biological and mechanical qualities that facilitate osseointegration. Compared to the original procedure, when metaphyseal bone defects are too severe, a single tantalum cone may not be enough and we have developed a technique that could extend the indications for this cone in these cases. We used 2 cones to fill femoral bone defects in 7 patients. There were no complications due to wear of the tantalum cones. Radiological follow-up did show any migration or loosening. The short-term results confirm the interest of porous tantalum cones and suggest that they can be an alternative to allografts or megaprostheses in case of massive bone defects.

Photoresponsive self-assemblies based on fatty acids.

Photoresponsive surfactant system based on fatty acids has been developed by the introduction in aqueous solution of a photoacid generator (PAG). Self-assembly transitions are triggered by UV irradiation due to a pH change induced by the presence of PAG.

Thrombopoietin induces histidine decarboxylase gene expression in c-mpl transfected UT7 cells.

The leukemic cell line UT7 is endowed with both megakaryocyte and basophil differentiation potential, as judged by its capacity to respond to PMA by displaying megakaryocytic and basophilic markers and to produce histamine by neosynthesis. Herein, we addressed the question whether the biological activities characteristic of basophil differentiation were still induced when c-mpl-transfected UT7 cells received a specific megakaryocytic differentiation signal delivered by thrombopoietin (TPO). Surprisingly, we found that histamine synthesis did effectively occur in response to the growth factor. This activity was not associated with megakaryopoiesis since it was not detected in megakaryocytes generated from CD34(+) cells cultured in the presence of TPO. Comparing different c-mpl-transfected cell lines, we found that the amount of histamine generated in response to TPO correlated with their responsiveness to PMA, but not with their level of c-mpl expression, thus revealing an intrinsic basophil differentiation potential. Both PMA- and TPO-induced histamine synthesis was reduced by PKC and MEKs inhibitors, indicating that the induction occurred through a common signalling pathway.

A distinct IL-18-induced pathway to fully activate NK T lymphocytes independently from TCR engagement.

NK T lymphocytes are characterized by their ability to promptly generate IL-4 and IFN-gamma upon TCR engagement. Here, we demonstrate that these cells can also be fully activated in the absence of TCR cross-linking in response to the proinflammatory cytokine IL-18 associated with IL-12. NK T cells stimulated with IL-18 plus IL-12 proliferated, killed Fas+ target cells, and produced high levels of IFN-gamma without IL-4. In these conditions, IFN-gamma production was at least 10-fold higher than that upon TCR cross-linking. Interestingly, a 2-h pretreatment with IL-12 plus IL-18 sufficed to maintain the high IFN-gamma-producing potential during subsequent stimulation with anti-TCR mAbs or with the specific Ag alpha-galactosylceramide. Similar effects were observed in vivo, because splenic CD4+ NK T cells from MHC class II-deficient mice secreted IFN-gamma without further stimulation when removed 2 h after a single injection of IL-12 plus IL-18. In conclusion, our evidence for activation of NK T lymphocytes in response to IL-18 plus IL-12 in the absence of TCR engagement together with the maintenance of preferential IFN-gamma vs IL-4 production upon subsequent exposure to specific Ags is consistent with the active participation of this cell population in innate as well as acquired cellular immune responses.

Increased fetal and extramedullary hematopoiesis in Fas-deficient C57BL/6-lpr/lpr mice.

In this study, we examined the consequences of Fas deficiency on hematopoiesis in C57BL/6-lpr/lpr mice. We found a striking extramedullary increase in hematopoietic progenitor cells, comprising erythroid and nonerythroid lineages alike. These modifications preceded the lymphadenopathy, because early progenitors (colony-forming units-spleen [CFU-S] day 8) were already augmented in day-18 fetal livers of the lpr phenotype. Three weeks after birth, CFU-S increased in peripheral blood and spleen and colony-forming cells (CFU-C) began to accumulate 1 to 3 weeks later. Extramedullary myelopoiesis augmented progressively in Fas-deficient mice, reaching a maximum within 6 months. By then, mature and immature myeloid cells had infiltrated the spleen, the liver, and the peritoneal cavity. Similar changes occurred in C57BL/6-gld/gld mice, indicating that they resulted from Fas/FasL interactions. Medullary hematopoiesis was not significantly modified in adult mice of either strain. Yet, the incidence of CFU-S decreased after Fas cross-linking on normal bone marrow cells in the presence of interferon gamma, consistent with a regulatory function of Fas/FasL interactions in early progenitor cell development. These data provide evidence that Fas deficiency can affect hematopoiesis both during adult and fetal life and that these modifications occur independently from other pathologies associated with the lpr phenotype.

Modulation of histidine decarboxylase activity and cytokine synthesis in human leukemic cell lines: relationship with basophilic and/or megakaryocytic differentiation.

In the present study, we show that UT7D1 cells, derived from the pluripotent cell line UT7, express high levels of histidine decarboxylase (HDC) mRNA spontaneously. These cells conserve the ability to differentiate into megakaryocytes upon stimulation with PMA, while greatly increasing their HDC activity. We provide evidence that enhanced HDC activity reflects the basophil rather than the megakaryocytic differentiation potential of UT7DI cells. Indeed, in addition to HDC mRNA, they express spontaneously several other mRNA coding for molecules present in basophils (FcepsilonRI, CCR3, IL-4Ralpha, IL-5Ralpha). Furthermore, the basophil antigen Bsp-1 is displayed on the surface of some UT7D1 cells in response to PMA concomitantly with increased histamine synthesis and mRNA expression of typical basophil-derived cytokines (IL-6, IL-4, and IL-13). Nevertheless, PMA cannot sustain the differentiation of this lineage, because mRNAs for basophil markers gradually diminish during long-term culture, whereas molecules associated with the megakaryocytic lineage remain prominent. In support of the notion that HDC activity is not related with megakaryopoiesis, we show that PMA-induced CD41 expression and PDGF transcription occurs in the K562 cells, though neither HDC mRNA nor any known basophil marker are expressed in these conditions. In contrast, all these markers are expressed in the basophilic leukemia cell line KU812F. Interestingly, the megakaryocytic cell line HEL produces also substantial amounts of histamine and expresses FcepsilonRI, thus revealing its basophil differentiation potential. HEL as well as KU812F need not be stimulated with PMA to react with Bsp-1 mAb, suggesting that they are more engaged into the basophil differentiation scheme than UT7D1. Other leukemic cell lines unrelated to the megakaryocyte or basophil lineage, like HL60 and U937 do neither synthesize histamine nor express basophil markers before or after PMA stimulation. To our knowledge, this is the first evidence for a factor-dependent cell line with megakaryocyte/basophil bipotentiality with which early stages of basophil commitment can be analyzed.

IL-3-induced coexpression of histidine decarboxylase, IL-4 and IL-6 mRNA by murine basophil precursors.

Murine low-density bone marrow cells sorted from the blast cell window on the basis of high rhodamine-123 retention (Rh-bright), are highly enriched in histamine-, IL-4-, and IL-6-producing cells. We established by in situ hybridization that up to 50% of this population (around 0.25% of the whole bone marrow) coexpressed the transcripts for these molecules upon stimulation with 1L-3. Rh-bright cells were also positive for mRNA encoding the alpha, beta, and gamma chains of the Fc(epsilon)RI which was functional since aggregated IgE induced the same percentage of cells hybridizing with the HDC probe as IL-3. Clonogenic progenitors and histamine- and cytokine-producing cells copurified in the Rh-bright population, but could be distinguished by their c-kit expression, CFU-C being more frequent in the c-kit(high) fraction, while histamine and IL-6 producers were enriched in the kit(low) counterpart. Ultrastructural analysis of Rh-bright cells revealed essentially two subsets, namely undifferentiated blast cells and basophil precursors. No other lineage-committed population was enriched by this sorting procedure, and it can therefore be concluded that coexpression of HDC, IL-6, and IL-4 transcripts in response to IL-3 or aggregated IgE takes place mainly in hematopoietic precursors belonging to the basophil lineage.

Prognostic value of vertebral lesions detected by magnetic resonance imaging in patients with stage I multiple myeloma.

We assessed the role of spinal magnetic resonance imaging (MRI) and bone densitometry as prognostic factors in patients with asymptomatic stage I multiple myeloma (MM) and negative skeletal survey. 55 consecutive patients underwent spinal MRI and 41 of them underwent bone densitometry by dual-energy X-ray absorptiometry (DEXA). Spinal MRI studies showed evidence of bone marrow involvement in 17/55 patients (31%). A diffuse pattern was present in three patients and a focal pattern in 14 patients, nine of them with only one nodular lesion. During a median follow-up of 25 months, 10 patients had disease progression, 8/17 patients with abnormal MRI and 2/38 patients with normal MRI. Median time to disease progression was not reached in both groups but was significantly different for patients with normal and those with abnormal patterns on MRI (P < 0.0001). Lumbar BMD was only slightly decreased compared with normal people (median lumbar Z score -0.43) and was not of prognostic value. Using a multivariate analysis the only two independent significant prognostic parameters were abnormal MRI (P<0.001, HR 30.4, 95% CI 4.3-213) and bone marrow plasmacytosis >20% (P=0.004, HR 16.4, 95% Cl 2.6-104). Thus, spinal MRI but not bone densitometry, appeared to be justified in patients with stage I asymptomatic MM and negative skeletal survey.

Fas cross-linking mimics the inhibitory effect of anti-CD3 on IL-3-induced histamine and cytokine production by murine myeloid spleen cell precursors.

In the present study we investigated the effect of anti-CD3 stimulation on IL-3-induced histamine, IL-6, and IL-4 synthesis by murine hematopoietic precursor cells. These activities were strikingly decreased in splenocytes from mice that had received a single intravenous injection of 10 microg of anti-CD3 monoclonal antibody (mAb) 24 hours previously. A similar inhibition occurred after 24-hour in vitro stimulation of normal spleen cells with 1 microg/mL of anti-CD3 mAb. In both situations the inhibitory effect depended on T cell activation in that treatment with F(ab')2 fragments of anti-CD3 did not diminish secretion of histamine and cytokines. Cross-linking of Fas antigen on spleen cells mimicked the action of anti-CD3, provided that interferon (IFN)-gamma was present during the incubation period. Substantial amounts of this cytokine were detected in spleen cell supernatants, which were able to replace recombinant IFN-gamma during Fas receptor cross-linking. This effect was entirely mediated by IFN-gamma, as assessed by its neutralization in the presence of anti-IFN-gamma mAbs. In contrast to splenocytes, bone marrow cells responded normally to IL-3 after in vivo or in vitro stimulation with anti-CD3. They were also not affected by combined treatment with anti-Fas mAb and IFN-gamma. Together, our data support the notion that the decrease in IL-3-induced histamine and IL-6 production by splenocytes pretreated with anti-CD3 is mediated, at least in part, by Fas/FasL interactions, suggesting that the activity of extramedullary myeloid precursor cells can be modulated by molecules involved in apoptosis.

IL-4-producing NK T cells are biased towards IFN-gamma production by IL-12. Influence of the microenvironment on the functional capacities of NK T cells.

NK T cells are an unusual T lymphocyte subset capable of promptly producing several cytokines after stimulation, in particular IL-4, thus suggesting their influence in Th2 lineage commitment. In this study we demonstrate that, according to the cytokines present in the microenvironment, NK T lymphocytes can preferentially produce either IL-4 or IFN-gamma. In agreement with our previous reports showing that their IL-4-producing capacity is strikingly dependent on IL-7, CD4-CD8-TCRalphabeta+ NK T lymphocytes, obtained after expansion with IL-1 plus granulocyte-macrophage colony-stimulating factor, produced almost undetectable amounts of IL-4 or IFN-gamma in response to TCR/CD3 cross-linking. However, the capacity of these T cells to produce IFN-gamma is strikingly enhanced when IL-12 is added either during their expansion or the anti-CD3 stimulation, while IL-4 secretion is always absent. A similar effect of IL-12 on IFN-gamma production was observed when NK T lymphocytes were obtained after expansion with IL-7. It is noteworthy that whatever cytokines are used for their expansion, IL-12 stimulation, in the absence of TCR/CD3 cross-linking, promotes consistent IFN-gamma secretion by NK T cells without detectable IL-4 production. Experiments in vivo demonstrated a significant upregulation of the capacity of NK T cells to produce IFN-gamma after anti-CD3 mAb injection when mice were previously treated with IL-12. In conclusion, we provide evidence that the functional capacities of NK T cells, which ultimately will determine their physiological roles, are strikingly dependent on the cytokines present in their microenvironment.

Hematopoietic progenitors and interleukin-3-dependent cell lines synthesize histamine in response to calcium ionophore.

The calcium ionophore A23187 promotes histamine synthesis in murine bone marrow cells by increasing the expression of mRNA encoding histidine decarboxylase (HDC), the histamine-forming enzyme. The cells responsible for this biological activity copurify with hematopoietic progenitors in terms of density, light scatter characteristics, and rhodamine retention, similar to interleukin (IL) 3-induced histamine-producing cells. Yet, the effect of calcium ionophore is not mediated by IL-3. The most purified rhodamine-bright bone marrow subset contains 80% cells that respond to calcium ionophore by increased HDC mRNA expression. This high frequency makes the involvement of one particular progenitor subset in histamine synthesis unlikely. The finding that all IL-3-dependent cell lines tested so far exhibit increased histamine production and HDC mRNA expression in response to calcium influx lends further support to this notion. Cell lines requiring other growth factors or proliferating spontaneously lack this ability. Finally, it should be noted that IL-3-dependent cell lines do not produce histamine in response to their growth factor. It might, therefore, be suggested that the pathway transducing the signal for increased histamine synthesis after IL-3 receptor binding in normal hematopoietic progenitors is modified in these cell lines.