Non-cryopreserved peripheral blood stem cells autologous transplantation in multiple myeloma: Bicentric study.

OBJECTIVES: The objective of this study is to evaluate the toxicity of autologous transplantation of non-frozen peripheral blood stem cells in Moroccan patients with multiple myeloma. MATERIAL AND METHODS: This was a bicentric retrospective study conducted in the Clinical Haematology Department of Mohammed V Military Teaching Hospital and at the Al Madina Clinic in Casablanca. The study period was from January 2015 to June 2019. All patients with multiple myeloma who had undergone an autologous peripheral stem cell transplant without freezing were included. Mobilisation was performed with lenograstim alone and the collected stem cells were stored for 24-48hours in a blood bank refrigerator at a temperature of 4°C. After standard conditioning with high-dose melphalan, the peripheral blood stem cells were reinjected 24 h following conditioning. RESULTS: Over the study period, 55 patients received an autologous transplant using non-frozen peripheral blood stem cells. The median richness of the CD34 cells collected was 4.5×106 CD34/kg (range: 2-12.2). The time required for neutrophil recovery was 12 days (range: 7-19). The time required for platelet recovery was 14 days (range: 9-32). The mortality rate within 100 days post-transplant was 3.6%. We did not observe any cases of graft failure. CONCLUSION: Our study finds good feasibility and low toxicity of autologous peripheral stem cell transplantation without freezing in patients with multiple myeloma.

HLA typing and haematopoietic stem cell transplantation in the histocompatibility unit of the Ibn Sina University Hospital in Rabat (Morocco).

PURPOSE OF STUDY: This study focuses on the search for a suitable related HLA-matched donor of haematopoietic stem cells in the context of allogeneic transplantation in Morocco. The aim of this work is to establish whether the related donor can meet graft needs in Moroccan patients. PATIENTS AND METHODS: 429 families (429 recipients and 965 donors) benefited from HLA typing, using microlymphocytotoxicity, polymerase chain reaction-sequence specific primer and/or high-resolution polymerase chain reaction-specific sequence oligonucleotide. RESULTS: The recipients and donors are mostly men over 18 years of age. In total, 86.8% of the recipients have between 1 and 3 donors who are 96% of the collaterals. Malignant haemopathies account for 54% of allograft indications. Benign haemopathies are more frequent than malignant in children, whereas the profile is reversed in adults. Fifty percent of recipients have an HLA identical donor in their siblings and 42% and HLA haplo identical donor. The HLA typing of the recipients and the donors reveals very large polymorphism of the population. CONCLUSION: The related donor of haematopoietic stem cells represents an important source of grafts but will not be able to satisfy all the needs of Morocco. The creation of national unrelated voluntary donors will open up new possibilities for recipients who do not have a compatible donor within his relatives.

[Recipients adverse reactions in the Ibn Sina Hospital of Rabat: State 1999-2013]. / Effets indésirables receveurs à l'hôpital Ibn Sina de Rabat : bilan 1999­2013.

PURPOSE OF STUDY: The declaration of the recipients adverse reactions (RAR) is one of the field haemovigilance activities. It provides an evaluation of transfusion side effects and thus prevents their appearance. The aim of this study is to analyze, over 14 years, the RAR supports reported in Rabat Ibn Sina hospital. PATIENTS AND METHODS: All of the RAR supports sending to the blood transfusion service were analyzed. The data collected from these supports are: clinical characteristics of the patient, type of incident observed and type of labile blood products (LBP) transfused. RESULTS: A total of 353 RAR were declared with a mean cumulative incidence of 1.7/1000 LBP delivered. Febrile non-hemolytic transfusion reactions represent 72.8% of the RAR declared. The RAR were classified as grade 1 in 87.1% of cases and were secondary to a transfusion of the red cell concentrates in 81.9%. ABO incompatibility was found in four cases (0.02/1000 LBP delivered). CONCLUSION: The number of RAR reported by Rabat Ibn Sina hospital remains underestimated. Management and traceability RAR and rigorous investigation, under the responsibility of the corresponding haemovigilance contribute to the improvement of transfusion safety.

[Immunological monitoring in kidney transplantation: 13 years experience of a Moroccan histocompatibility laboratory]. / Suivi immunologique en transplantation rénale : 13 ans d'expérience d'un laboratoire d'histocompatibilité marocain.

PURPOSE OF STUDY: The quality of the immunological monitoring is crucial because it determines the success of the kidney transplantation. The scope of this work is to describe the experience of the department of immunological unity of the Ibn Sina university hospital in Rabat regarding the immunological monitoring of patients transplanted between 2001 and 2014. PATIENTS AND METHODS: Patient samples were collected from nephrology services of different public and private hospitals of Morocco. The tests conducted in the context of immunological monitoring are ABO typing, HLA-A, B, DR, DQ typing, anti-HLA antibodies detection and identification and cross-match. RESULTS: One hundred and fourteen benefited from a pre- and post-transplant immunological monitoring in our laboratory. The percentage of recipients having between 2 and 5 stored sera is 60.5 before transplantation and 56.1 after transplantation. Immunized patients account for 22.8% before the transplant and 17.6% after transplantation. Ninety-seven patients still have a functional graft, while 4 of them had DSA of low intensity before transplantation. Five immunological rejections were reported while the cross-match were negative and no DSA was identified before transplantation. Patient survival and graft at 1 year was 98.2% and 92.7% respectively. CONCLUSION: Conducting regular immunological monitoring is sometimes difficult in our context, however, the results are satisfactory in terms of graft and patients survival.

[The HLA system in the Moroccan population: General review]. / Le système HLA dans la population Marocaine: revue générale.

The Moroccan population is an interesting study model of Human Leukocyte Antigen (HLA) polymorphism given its ethnic and genetic diversity. Through an analysis of the literature, this work proposes to establish a balance of knowledge for this population in the field of histocompatibility: HLA diversity, anthropology, transplantation and HLA associations and diseases. This analysis shows that the HLA system has not been fully explored within the Moroccan population. However, the results obtained allowed us to initiate a database reflecting the specific healthy Moroccan population HLA polymorphism to identify immigration flows and relationships with different people of the world and to reveal the association of certain HLA alleles with frequent pathologies. We also propose to analyze the reasons hindering the development of this activity in Morocco and we will try to identify some perspectives.

Family-based association study of HLA class II with type 1 diabetes in Moroccans.

BACKGROUND: The T1D is a multifactorial disease; with a strong genetic control. The human leukocyte antigen (HLA) system plays a crucial role in the autoimmune process leading to childhood diabetes. About 440,000 of the childhood population of the world (1.8 billion children under 14 years of age), have type 1 diabetes, and each year an additional 70,000 develop this disorder. The objective of this study was to investigate the distribution of HLA class II in Moroccan families of diabetic children to identify susceptibility alleles of the Moroccan population. SUBJECTS AND METHODS: We included in this study, Moroccan families who have at least one child with T1D. The age of onset of diabetes was less than 15 years. HLA class II (DRB1* and DQB1*) was carried out by molecular biology techniques (PCR-SSP and PCR-SSO). The FBAT test (family-based association test) was used to highlight the association between T1D and the HLA-DRB1* and -DQB1* polymorphism. RESULTS: The association of HLA class II (DRB1*, DQB1*) in type 1 diabetes was analyzed in fifty-one Moroccan families, including 90 diabetics. The results revealed that the most susceptible haplotypes are the DRB1*03:01-DQB1*02:01, DRB1*04:05-DQB1*03:02 (Z=3.674, P=0.000239; Z=2.828, P=0.004678, respectively). And the most protective haplotype is the DRB1*15-DQB1*06. CONCLUSION: This is the first family-based association study searching for an association between HLA class II and T1D in a Moroccan population. Despite the different ethnic groups forming Morocco, Moroccan diabetics share the most susceptible and protective HLA haplotypes with other Caucasians populations, specifically the European and Mediterranean populations.

Genetic factors and multiple sclerosis in the Moroccan population: a role for HLA class II.

BACKGROUND AND OBJECTIVE: Multiple sclerosis (MS) is an autoimmune inflammatory demyelinating disease of the central nervous system that mainly affects young adults. The association between susceptibility to MS and HLA class II genes, in particular the DRB1*15 allele, has been reported in diverse ethnic groups. The aim of our study was to investigate the distribution of HLA-DRB1* and -DQB1* alleles in Moroccan population and their implication in the susceptibility to the disease. METHODS: Fifty-seven MS patients were compared to 172 healthy controls unrelated to one another and matched by age, sex and ethnic origin. HLA class II (DRB1* and DQB1*) typing was performed by PCR-SSP and/or Luminex (PCR-SSO). Allelic and haplotypic frequencies, P-values, odds ratio (OR) and 95% confidence interval (CI) were calculated using the software SPSS. RESULTS: A significant increase of DRB1*15 allele frequency (17.6% vs 8.4%, OR=2.67, 95% CI=1.36-5.23, P=0.004) and HLA-DRB1*15-DQB1*06 haplotype (8.8% vs 4.08%, OR=2.78, 95% CI=1.41-5.48, P=0.002) were observed in Moroccan MS patients. No association of the DR15 allele with sex or age at onset was appreciated. Concerning HLA-DQB1* alleles, no significant difference between patients and controls was found. CONCLUSIONS: Our results reveal a role for HLA-DRB1*15 allele molecules in the predisposition of Moroccan patients to MS. Although this study should be confirmed on a larger sample size, it analyzes for the first time the possible role of a genetic marker for susceptibility to MS in Moroccan population.

[Management by objectives: an experience by transfusion and immunology service in Rabat]. / Contractualisation interne : expérience du service de transfusion et d'immunologie de Rabat.

The management by objectives method has become highly used in health management. In this context, the blood transfusion and haemovigilance service has been chosen for a pilot study by the Head Department of the Ibn Sina Hospital in Rabat. This study was conducted from 2009 to 2011, in four steps. The first one consisted in preparing human resources (information and training), identifying the strengths and weaknesses of the service and the identification and classification of the service's users. The second step was the elaboration of the terms of the contract, which helped to determine two main strategic objectives: to strengthen the activities of the service and move towards the "status of reference." Each strategic objective had been declined in operational objectives, then in actions and the means required for the implementation of each action. The third step was the implementation of each action (service, head department) so as to comply with the terms of the contract as well as to meet the deadlines. Based on assessment committees, the last step consisted in the evaluation process. This evaluation was performed using monitoring indicators and showed that management by objectives enabled the Service to reach the "clinical governance level", to optimize its human and financial resources and to reach the level of "national laboratory of reference in histocompatibility". The scope of this paper is to describe the four steps of this pilot study and to explain the usefulness of the management by objectives method in health management.

Association of HLA alleles and haplotypes with vitiligo in Moroccan patients: a case-control study.

The aim of this study was to identify HLA class II alleles that may be involved in vitiligo genetic susceptibility in the Moroccan population and to determine susceptible and protective HLA alleles/haplotypes in vitiligo. One-hundred unrelated vitiligo patients and 300 healthy unrelated controls were studied for HLA class II alleles by polymerase chain reaction-sequence-specific primers. The phenotypic frequency of DRB1*07 (OR = 2.23, p c = 0.014) was significantly higher, while that of DRB1*03 (OR = 0.40, p c = 0.014) was significantly lower in patients than in controls. Haplotype DRB1*07-DQB1*02 (OR = 2.25, p c = 0.024) was positively associated with vitiligo patients, while haplotype DRB1*03-DQB1*02 (OR = 0.35, p c = 0.012) was negatively associated with this group. Vitiligo patients with positive family history and negative anti-thyroid peroxidase antibody (anti-TPO) have an extremely high phenotype frequency of DRB1*07-DQB1*02 haplotype (OR = 2.91, p c = 0.048 and OR = 2.62, p c = 0.00475, respectively). DRB1*03-DQB1*02 (OR = 0.32, p c = 0.048 and OR = 0.38, p c = 0.048, respectively) was negatively associated with patients without a family history and negative anti-TPO. This study demonstrated the positive association of HLA class II alleles and haplotypes with vitiligo in the Moroccan population. There may be differences in HLA haplotypes distribution in patients according to family history and anti-TPO profile.

Distribution of HLA class I and II genes in ankylosing spondylitis patients from Morocco.

OBJECTIVES: In Morocco, the patients affected by ankylosing spondylitis (AS) presents a high frequency of coxitis. Our study reports, for the first time, the polymorphism of Human Leukocyte Antigen (HLA) class I and class II molecules in the Moroccan patients. METHODS: Forty-six patients diagnosed with an AS and coxitis were compared to a group of 183 healthy controls matched by age, sex and ethnic origin. The HLA typing was performed using microlymphocytotoxicity for the class I (-A, -B) and PCR-SSP for the class II (-DR, -DQ). RESULTS: We found a significant increase of the HLA-B27 antigen frequency (P<0.0001, RR=20.9) in AS patients (29.3%) compared to the controls (3.2%) and a significant decrease in the frequency of HLA-B12 and HLA-B18 antigens. Examination of HLA class II distribution shows a significant increase of the HLA-DRB1*11 allele frequency in patients (P<0.0001). Concerning HLA-DQB1* alleles, no significant difference between patients and controls was appreciable. CONCLUSIONS: The HLA-B27 antigen is involved in the predisposition to the AS with coxitis in the Moroccan population. However, the low frequency observed in our population suggests the existence of other genetic and/or environmental factors. Other HLA genes seem to confer a predisposing effect (DRB*11) or a protective effect (B12 and B18) against the disease.

[Traceability of labile blood products in Morocco: experience of the Ibn-Sina hospital of Rabat between 1999 and 2010]. / Traçabilité des produits sanguins labiles au Maroc : expérience de l'hôpital Ibn-Sina de Rabat entre 1999 et 2010.

PURPOSE OF STUDY: The blood transfusion and haemovigilance service of the Ibn-Sina hospital in Rabat (Morocco) was created 1997. This unit manages the pretransfusional tests, distribution of blood products, traceability and haemovigilance. The objective of this study was to analyze, over a period of 12years, the traceability of blood products delivered in our hospital and the measures used to improve feedback information. PATIENTS AND METHODS: This is a retrospective study conducted between 1999 and 2010. Traceability rate was calculated from the feedback of traceability forms supplied with blood products (number of blood products noted on traceability forms on the total number of delivered product). To improve traceability rate, several actions were undertaken: one-time training, awareness campaigns and call phones asking for feedback information. RESULTS: Between 1999 and 2010, the service has delivered 173,858 blood products. The average rate of traceability during this period was 13.4 %. Traceability rate varies widely over time (5.2 % in 1999, 15.5 % in 2010) and shows a maximum value of 27.2 % in 2005. Feedback information is lower in emergency departments than in medical and surgical services. CONCLUSION: Feedback information about traceability in Ibn-Sina hospital remains very poor despite the measures used. Other actions, such as continuous education courses, low enforcement and informatisation should be considered.

[The latest TH17 lymphocyte in the family of T CD4+ lymphocytes]. / Le lymphocyte TH17 dernier-né de la famille des lymphocytes T CD4+.

In recent years the T CD4+ lymphocyte family has grown. In the initial two components TH1 and TH2 lymphocytes were added the TH17 lymphocyte and T cell regulator (Treg). Under the influence of transforming growth factor ß, interleukin 6 (IL6), IL21 and IL23, the naive lymphocyte T CD4+ differentiates in TH17. Currently, the TH17 is recognized as the leading actor of local inflammation through the pro-inflammatory cytokines (interleukins 17, 21, 22) that secretes and the expansion and recruitment of neutrophils that leads. Therefore, it is involved in chronic inflammatory processes, autoimmune diseases (rheumatoid arthritis, systemic lupus erythematosus), allergy and rejection of allogeneic transplants. TH17 lymphocyte opens up new therapeutic prospects for these pathologies.

[Toll-like receptors]. / Toll-like récepteurs.

The toll-like receptors are innate immunity receptors which recognise particular exogenous structures in the microorganisms pathogen associated molecular pattern (PAMP) and endogenous structures damage-associated molecular patterns (DAMP). Eleven TLR have been identified among human beings. These are danger receptors located in the cells of the immune system but also in other cells. Their primary function is the recognition of pathogens and the activation of the cell that holds them. It follows from it an action on the cells environment, inflammation cells and an activation of the adaptive immunity. The knowledge of the intracellular signalisation ways of the TLR has allowed us to understand the physiopathology of certain diseases. Thus, several works use the agonists of TLR to stimulate them: vaccines against infectious diseases, allergies and cancers. The antagonists are used to block the TLR in autoimmune and chronic inflammatory diseases. It is clear that the border between innate and adaptive immunity fades and that these two components of the immune response are closely related, thus opening up new prospects diagnostic and therapeutic procedures.

[HLA polymorphism and Behçet's disease in Moroccan population]. / Polymorphisme HLA et maladie de Behçet dans la population marocaine.

OBJECTIVES: We have study the various associations between Behçet disease and alleles HLA class I and II in Moroccan population. The distribution of HLA alleles involved in the disease is assessed according to clinical signs and sex. PATIENTS AND METHODS: Patients suffering from Behçet's disease have been compared to healthy controls matched by age and ethnic origin. The HLA typing has been conducted by using microlymphocytotoxicity for the class I (A and B) and molecular biology (polymerase chain reaction-sequence specific primers) for the class II (DR and DQ). RESULTS: In addition to the allele B51, -A2, -B5102, -B58 and -B72 predispose to the Behçet's disease and the alleles A23, -A33, -B18, -B41 and -B49 have a negative correlation in our Moroccan patients. The alleles A2 and -B72 are specific to men, -A68 and -B58 to women, whereas -B51 and -B5102 are predisposing to the Behçet's disease of both genders. The allele B51 is related to the mucous manifestations, -B72 to the vascularitis manifestations and -B58 to the cutaneous manifestations. The haplotypes A2-B51 has a high positive correlation with this Behçet's disease. Concerning class II, none of the alleles DRB1* is implicated in the pathology even though the allele DQB1*02 ensures the patients' protection. CONCLUSIONS: The allele HLA-B51 shows a high positive correlation with the Behçet's disease. Other genetic factors seem to be implicated in susceptibility or protection against the disease.

Pemphigus and HLA in Morocco.

PURPOSE OF STUDY: Pemphigus is a group of autoimmune bullous dermatosis diseases characterized by autoantibodies against keratinocyte adhesion molecule. A significant association with HLA class II genes, particularly DR4 and DR14 has been described in many ethnic groups and countries. We have investigated, for the first time in Morocco the relationship between different pemphigus subtypes and HLA genes. PATIENTS AND METHODS: Fifty-two unrelated patients were compared to 178 healthy controls matched by age, sex and ethnic origin. HLA typing was performed by standard complement dependent microlymphocytotoxic method for class I and by sequence-specific primer amplification method for class II. RESULTS: No significant association was observed with any of the HLA-A or -B antigens. Generic typing showed a significant increase of DRB1*04 (p=0.002), DRB1*14 (p=0.003) and DQB1*03 (p=0.02) allele frequencies and significant decrease of DRB1*15 (p<0.0001) and DQB1*06 (p=0.01) allele frequencies. HLA-DRB1*15-DQB1*06 haplotype seems to confer a protective effect in our population while DRB1*04-DQB1*03 and DRB1*14-DQB1*05 haplotypes induced susceptibility to the disease. CONCLUSION: Taken together, our results confirmed the genetic predisposition to pemphigus. However, genetic factors are not sufficient to explain the high prevalence of pemphigus observed in the Moroccan population since alleles of susceptibility were similar to those commonly described in other populations throughout the world.

HLA-A, -B, -DR and -DQ allele and haplotype frequencies in the Moroccan population: a general population study.

PURPOSE OF STUDY: The aim of this general population study is to extend our knowledge about the HLA-A, -B, -DR and -DQ genes distribution and their diversity among the Moroccan population. PATIENTS AND METHODS: One hundred and ten unrelated healthy Moroccans from diverse regions of the country were included in the study. HLA typing was done either by serological (standard complement dependent microlymphocytotoxicity) and/or molecular (sequence-specific primer amplification) techniques. RESULTS: The most frequent alleles observed were: HLA-A2 (19.1%), -A1 (10%), -A3 (10%), -B44 (9.6%), -DR3 (17.1%), -DR4 (15.3%), -DQ2 (30.6%), -DQ6 (26.4%) and -DQ3 (25%). No predominant haplotype was observed for HLA A-B while high frequency was observed for some HLA DR-DQ associations (DR3-DQ2, DR15-DQ6, DR7-DQ2, DR4-DQ3, DR13-DQ6). Comparison with neighbouring populations, on the basis of alleles, haplotypes and genetic distances, showed that Moroccan population is close to the Algerians, the Tunisians, the Spaniards and the French. The haplotype frequencies revealed also relationships with Italians, Sardinians, Basques, Portuguese and Moroccan Jews populations. CONCLUSION: Our results confirm and extend the current knowledge about the Moroccan genetic pattern and reflect all the ethnic diversity of the country. This study will be helpful in the future for clinical analysis like transplantation and HLA-associated diseases in Moroccan population.