Clinicopathological Study of Males with Lupus Nephritis: Pathologist's Experience at a Tertiary-Care Center.

Background: Systemic lupus erythematosus (SLE) is an autoimmune systemic disorder, more common in females of reproductive age-group as compared with males. There are very few studies regarding lupus nephritis (LN) in males. Hence, we decided to study the clinical and pathological findings of LN in males. Materials and Methods: We carried out a retrospective study over a period of 5 years (January 2014-December 2018) on indicated native renal biopsies from male patients with LN. We analyzed the clinical, laboratory, and histological findings of these patients. Results: Renal biopsies were performed on 228 patients with LN, of which 29 (12.72%) biopsies were in male patients. The mean age at presentation was 28.3 ± 12.98 years. Edema (65.5%) was the most common clinical feature followed by arthritis (27.58%), fever (27.58%), and skin rash (24.1%). The mean values for 24 hours urinary protein, serum double-stranded DNA, serum antinuclear antibody, and serum complement C3 were 4.98 ± 2.91 g, 137.7 ± 91.93 IU/mL, 2.96 ± 1.78, and 65.07 ± 36.30 mg/dL, respectively. On histology, the most common class of LN was Class IV (34.48%) followed by Class V (20.68%), combined Class IV + V (20.68%), Classes II, III, and III + V. Conclusion: LN can affect males, although the prevalence is lower than in females. The incidence of LN in our study was 12.7% with the most common histological class being diffuse proliferative LN.

Anti-phospholipase A2 Receptor Antibody in Differentiation and Prognostication of Membranous Nephropathy.

Anti-M-type phospholipase A2 receptor (anti-PLA2R) antibody is believed to be associated with primary membranous nephropathy (pMN) and absent in secondary MN (sMN). There are few data regarding utility of anti-PLA2R antibody as a prognosticator. Our study aimed to compare the incidence of positive serum anti-PLA2R antibody titer in pMN versus sMN and correlation with clinical outcome. From August 2015 to July 2019, patients with biopsy-proven MN were evaluated for serum anti-PLA2R antibody titers by the enzyme-linked immunosorbent assay. The subset of cases was repeated to monitor the clinical response in terms of 24 h proteinuria. A total of 169 patients, 65 pMN and 104 sMN were studied. Anti-PLA2R antibody was found in 41 (63.08%) pMN with mean titer, 232.62 RU/mL, and 40 (38.46%) sMN with mean titer 253.59 RU/mL. Out of positive antiPLA2R antibody titer in pMN cases, 15 were retested twice to 5 times with mean titers of 78.95, 36.27, 13.9, and 15.45 RU/mL, respectively. Out of positive anti-PLA2R antibody in sMN cases, 11 were retested twice to five times with mean titers of 104.42, 122.49, 12.33, and 17.2 RU/mL, respectively. All patients with decreasing anti-PLA2R antibody titer in both groups had clinical remission, with a decrease in mean 24 h proteinuria from 7.11 g to 3.36 g in pMN and 5.97 g to 3.41 g in sMN. Ten pMN and 11 sMN patients without remission showed persistent positive anti- PLA2R antibody titer. Anti-PLA2R antibody titer may be elevated in pMN/sMN. It can also be used as a noninvasive prognostic marker for MN.

Ten-year appraisal of pediatric renal allograft biopsies: Points to ponder.

There is paucity of literature on pediatric renal allograft biopsy (RAB) evaluation. We present RAB findings of pediatric renal transplantation (RT) and correlate with outcome. This is a 10-year retrospective study of diagnostic RAB of children <12 years divided in to three groups: Group 1 (n = 9): less than haplo-match living donor RT (LDRT), Group 2 (n = 32): greater than or equal to haplo-match LDRT, and Group 3 (n = 7): deceased donor RT. Demographics, biopsy findings, survival, and serum creatinine (SCr) were evaluated. Statistical analysis was performed using IBM SPSS Statistics version 20.0. The most common findings were antibody-mediated rejection (ABMR) observed in 77.7%, 45%, and 71.5% and T-cell-mediated rejections (TCMRs) in 33.3%, 52.5%, and 42.9% in Groups 1, 2, and 3, respectively. Recurrent oxalosis was seen in 5% in Group 2. Death-censored graft survival was 100% at 1 year and 43.8% from 5 to 9 years in Group 1; 93.5%, 76.6%, 56.5%, and 14.4% at 1, 5, 10, and 15 years in Group 2; 100% at one year; and 71.4% from 5 to 12 years in Group 3. No patient appeared after 9 years in Group 1 and after 12 years in Group 3. In Group 1, the mean SCr (mg/dL) was 1.06 ± 0.45, 2.12 ± 1.87, and 1.39 at 1, 5, and 9 years; 1.35 ± 0.97, 1.73 ± 1.15, and 2.49 ± 1.64 in Group 2; and 1.15 ± 1.24, 1.43 ± 0.1, and 1.18 ± 0.06, respectively, in Group 3 at 1, 5, and 10 years posttransplant. ABMR followed by TCMR was the most common injury in all the groups. Group 1 had more rejections than others.

Thrombotic microangiopathy in a renal allograft: Single-center five-year experience.

Thrombotic microangiopathy (TMA) is devastating for renal transplantation (RT) causing graft/ patient loss. We present 5-year experience of TMA in RT in retrospective study of indicated renal allograft biopsies with TMA. Patient-donor demographics and associated histological findings with respect to transplants under tolerance induction protocol (Group 1) were compared with patients transplanted under triple immunosuppression (Group 2). Statistical analysis was performed using IBM SPSS Statistics version 20. Sixty-one (4.1%) of 1520 biopsies [Group 1:17 (1.9%)/882, Group 2:44 (6.9%)/638] revealed TMA. Tacrolimus trough levels were normal. There was no evidence of systemic involvement in any patient. Mean age was 36.8 years with 70.6% males, HLA-match, 2.6/6, and the most common original disease unknown (41.2%) in Group 1, and 35.9 years with 86.4% males, HLA-match, 2.1/6, and the most common original disease unknown (50%) in Group 2. Biopsies were performed at mean 5.1-year posttransplant in Group 1 and 2.3 years in Group 2. Acute TMA constituted 47% Group 1 and 43.2% Group 2 biopsies; of these, antibody-mediated rejections were observed in 58.8%, T-cell mediated rejections in 11.8%, tacrolimus toxicity in 76.5%, and other findings in 35.3% Group 1; and 61.4%, 25%, 50%, and 18.2%, respectively, in Group 2 biopsies. Higher rejection activity scores were more in Group 2. Postbiopsy 1- and 5- year patient survival was 94.1%, 86.9% in Group 1 and 92.1%, 88.3% in Group 2; 1- and 4-year graft survival was 52.9%, 15.9% in Group 1 and 20.3%, 5.4% in Group 2. TMA was poor prognosticator for RT, especially under triple immunosuppression. Antibody- mediated rejection and tacrolimus toxicity were more prone to TMA.

In vitro differentiation of neural cells from human adipose tissue derived stromal cells.

BACKGROUND: Stem cells, including neural stem cells (NSCs), are endowed with self-renewal capability and hence hold great opportunity for the institution of replacement/protective therapy. We propose a method for in vitro generation of stromal cells from human adipose tissue and their differentiation into neural cells. MATERIALS AND METHODS: Ten grams of donor adipose tissue was surgically resected from the abdominal wall of the human donor after the participants' informed consents. The resected adipose tissue was minced and incubated for 1 hour in the presence of an enzyme (collagenase-type I) at 370C followed by its centrifugation. After centrifugation, the supernatant and pellets were separated and cultured in a medium for proliferation at 370C with 5% CO2 for 9-10 days in separate tissue culture dishes for generation of mesenchymal stromal cells (MSC). At the end of the culture, MSC were harvested and analyzed. The harvested MSC were subjected for further culture for their differentiation into neural cells for 5-7 days using differentiation medium mainly comprising of neurobasal medium. At the end of the procedure, culture cells were isolated and studied for expression of transcriptional factor proteins: orthodenticle homolog-2 (OTX-2), beta-III-tubulin (ß3-Tubulin), glial-fibrillary acid protein (GFAP) and synaptophysin-ß2. RESULTS: In total, 50 neural cells-lines were generated. In vitro generated MSC differentiated neural cells' mean quantum was 5.4 ± 6.9 ml with the mean cell count being, 5.27 ± 2.65 × 103/µl. All of them showed the presence of OTX-2, ß3-Tubulin, GFAP, synaptophysin-ß2. CONCLUSION: Neural cells can be differentiated in vitro from MSC safely and effectively. In vitro generated neural cells represent a potential therapy for recovery from spinal cord injuries and neurodegenerative disease.

C4d-negative antibody-mediated rejection: A pathologist's perspective and clinical outcome.

Banff'13 update included C4d-antibody-mediated rejection (ABMR) as a separate entity responsible for graft dysfunction with limited clinical/prognostic implications. We present a retrospective study to determine the incidence and outcome of C4d-negative ABMR. A total of 987 renal allograft (RA) biopsies obtained from 987 RA recipients were studied from January 2013 to January 2016. All samples were subjected to light microscopy using standard stains and C4d immunohistochemistry on paraffin sections and reported according to modified Banff's criteria. Adequate biopsies with immunological injuries were categorized as Group 1: pure ABMR, Group 2: combined ABMR with concurrent T-cell-mediated rejection (TCR), and Group 3: pure TCR. Groups 1 and 2 were further subgrouped as C4d positive (Group 1a and 2a) or C4d negative (Group 1b and 2b). Graft function was measured by serum creatinine (SCr) level (mg/dL). Of the 987 biopsies, 43.3% (404) biopsies revealed immunological injury. Of these, 27.7% of the biopsies revealed pure ABMR (Group 1), 60.6% revealed combined ABMR with TCR (Group 2), and 11.3% revealed pure TCR (Group 3). The overall incidence of ABMR (pure ABMR + ABMR with TCR) was 36.27%, of which C4d-negative rejections were 18.48% and 18.7% in Group 1 and Group 2, respectively. The mean SCr at the end of three years follow-up in patients with C4d-negative rejections was comparatively higher. C4d-negative ABMR, recently included in Banff'13, has a low incidence, usually presents early after transplantation but carries better outcome than C4d-positive ABMR. However, further long-term studies are still required for knowing the clinical course over years.

Urinary Screening for Early Detection of Kidney Diseases.

OBJECTIVE: Urinary screening is a simple inexpensive tool to evaluate kidney functions. The authors carried out urinary screening of school children for early detection of kidney diseases. METHODS: Children in the age group 5-15 y were screened for urinalysis. They were divided in 2 groups; group-1 included 5-10 y and group-2 included >10-15 y old children. RESULTS: Urine samples of 3340(78%) out of 4283 enrolled children were tested. Abnormal samples were found in 5.75%; with proteinuria in 4.59%, pyuria in 3.29% and hematuria in 4.31%. Males constituted 47.71% in group-1 and 54.64% in group-2. Low body mass index was found in 94.1% group-1 and 78.99% group-2 children. Mild proteinuria was found in 1.2% group-1 and 2.56% group-2 children. Severe proteinuria was more in group-2 (0.77% vs. 0.06%) with female preponderance. Glucosuria was found in 1 boy of group-2. Urobilinogen was more in group-2 (0.65% vs. 0.24%) with male preponderance. Nitrituria was found in 9 girls. Pyuria (2.02% vs. 1.27%) and hematuria were more in group-2 (3.04% vs. 1.87%) with female preponderance. Combined proteinuria and hematuria (0.42% vs. 0.24%) as well bacteruria and fungaluria were more in group-2 (4.11% vs. 1.39%). Six of 192 children with abnormal urinary findings were treated; 1 for urinary calculus and 5 for urinary tract infection. CONCLUSIONS: Abnormal urinary findings were more common in children >10 y of age. Thus urinary screening program of children can become useful for early detection of kidney diseases and contribute towards building up of a healthy nation.

Primary Tubulovillous Adenocarcinoma of Renal Pelvis: An Unusual Genitourinary Tumor.

Prolonged inflammation or irritation due to renal calculi can induce glandular metaplasia of the urothelium and even malignant neoplasm. Primary adenocarcinoma in pelvicalyceal system is a rare tumor in such patients. Here, a case of carcinoembryonic antigen (CEA) positive primary tubulovillous adenocarcinoma in pelvicalyceal system is reported. A 57-year-old male with right abdominal pain and microscopic hematuria and the history of nephrolithotomy for recurrent renal calculi referred to our center. Radiological findings showed well-defined lobulated mass with calcification in interpolar region of right kidney extending up to pelvicalyceal system along with calculi in pelvicalyceal system and ureter with moderate hydroureteronephrosis. Laparoscopic right radical nephrectomy was performed. Histopathological examination revealed tubulovillous adenocarcinoma of renal pelvicalyceal system with CEA positivity. Patient was asymptomatic and had no recurrence after one and a half years. Primary tubulovillous adenocarcinoma in renal pelvis though rare, is usually associated with intestinal metaplasia of urothelium induced by prolonged chronic inflammation and renal calculi.

Six years' experience of tolerance induction in renal transplantation using stem cell therapy.

Tolerance induction (TI) has been attempted with chimerism/clonal deletion. We report results of TI protocol (TIP) using stem cell therapy (SCT) included adipose derived mesenchymal stem cells (AD-MSC) and hematopoietic stem cells (HSC) in 10 living-donor related renal transplantation (LDRT) patients under non-myeloablative conditioning with Bortezomib, Methylprednisone, rabbit-anti-thymoglobulin and Rituximab, without using conventional immunosuppression. Transplantation was performed following acceptable lymphocyte cross-match, flow cross-match, single antigen assay and negative mixed lymphocyte reaction (MLR). Monitoring included serum creatinine (SCr), donor specific antibodies (DSA) and MLR. Protocol biopsies were planned after 100days and yearly in willing patients. Rescue immunosuppression was planned for rejection/DSA/positive MLR. Over mean 6±0.37year follow-up patient survival was 80% and death-censored graft survival was 90%. Mean SCr was 1.44±0.41mg/dL. This is the first clinical report of sustained TI in LDRT for 6years using SCT.

Clinicopathological study of nondiabetic renal disease in type 2 diabetic patients: A single center experience from India.

Diabetic nephropathy (DN) is a major complication of diabetes mellitus (DM), leading to chronic kidney disease/end-stage renal disease. Wide spectrum of nondiabetic renal diseases (NDRD) is reported in type-2 diabetes (type-2 DM). We carried out this single-center study to find clinical, laboratory, and histological features of NDRD in type-2 DM patients and to assess the prevalence of NDRD in India. A single-center retrospective study which included analysis of renal biopsies from patients with type-2 DM, performed between January 2008 and September 2016. Biopsy findings were categorized into three groups, Group-I (isolated NDRD); Group-II (NDRD superimposed on underlying DN); and Group-III (isolated DN). Out of 152 diabetic patients (111 males and 41 females), 35 (23.03%) patients were of Group-I (isolated NDRD), 35 (23.03%) of Group-II (NDRD superimposed on underlying DN), and 82 (53.95%) of Group-III (isolated DN). The mean age (in years) was 55.08 ± 10.71, 55.65 ± 8.71, and 54.45 ± 9.01 respectively in Group-I, II, and III. Nephrotic syndrome (NS) was the most common clinical presentation in all groups. Duration of DM was significantly shorter in Group-I than in Group-II. Diabetic retinopathy was absent in Group-I. Proteinuria was more in Group-III than Group-I. Low serum C3 and/or C4 levels was observed in five (14.29%) cases of Group-I and Group-II each and two (2.43%) cases of Group-III. Nearly, 70 (46.05%) patients were found to have NDRD either in isolated form or as combined lesions. The most common histological types of NDRD were acute tubulointerstitial nephritis (38.57%) followed by benign nephrosclerosis (15.72%), membranous nephropathy (10%), IgA nephropathy (7.14%), and membranoproliferative glomerulonephritis (7.14%). The incidence of NDRD (with/without DN) in type-2 DM is very high. Shorter duration of diabetes, hematuria, absence of retinopathy, low serum complement levels, and nephrotic range proteinuria are predictors of NDRD.

Atypical Presentation of Isolated Cerebral Aspergillosis in a Renal Allograft Recipient.

The immunosuppressive state in organ transplantation leads to infectious complications responsible for high mortality rate. Fungal infections account for 5% of all infections in Renal Transplantation (RT) recipients. Aspergillus species are filamentous fungi frequently causing fungal infections in RT recipients. Lungs and paranasal sinuses are the usual portal of entry from where it disseminates to other organs. Here, we are reporting a case of 14-year-old boy with RT from mother's kidney, who had atypical presentation of isolated cerebral aspergillosis at 19 months post-transplant without identified portal of entry. Early diagnosis and prompt treatment saved the patient and the graft.

Repercussions of eosinophils in a renal allograft - Predictor of early graft loss!

We present 5-year experience of renal transplantation (RT) with tissue eosinophilia (TE) in renal allograft biopsy (RAB) and its repercussions on the outcome. In total, 1217 recipients underwent RT from 2011 to 2015, and they were evaluated for the presence of ≥4% TE. Group 1 consisted of RT with RAB showing TE, Group 2 consisted of RT with RAB with rejections without TE, and Group 3 consisted of RT without rejections. Group 1 had 27 recipients, Group 2 had 395, and Group 3 had 795 recipients. The outcome in terms of graft function, patient and graft survival were evaluated and compared between three groups. All recipients received standard triple immunosuppression. One-year patient and death-censored graft survival were 80.7% and 82.7% in Group 1, 87.2% and 95.1% in Group 2, and 92.6% and 99.6%, respectively in Group 3 and corresponding mean serum creatinine (SCr, mg/dL) was 1.60 ± 0.45 in Group 1, 1.63 ± 0.58 in Group 2, and 1.19 ± 0.39 Group three, respectively. Five-year patient and death-censored graft survival were 72.9 % and 71.1% for Group 2 and 87% and 98.2% for Group 3 with SCr of 1.63 ± 0.38 and 1.25 ± 0.4, respectively. Group 1 recipients did not appear at five years. At four years posttransplant, patient and death-censored graft survival were 71.7% and 59.5% in Group 1 with SCr of 1.55 ± 0.65 mg/dL. In conclusion, the presence of eosino-phils in a renal allograft is an impending sign of graft damage and eventual graft loss.

De Novo Focal Segmental Glomerulosclerosis in Renal Allograft-Histological Presentation and Clinical Correlation: Single Centre Experience.

INTRODUCTION: Recurrent or de novo glomerulonephritis are one of the well-known causes for renal allograft dysfunction in early and late period after renal transplantation. Focal Segmental Glomerulosclerosis (FSGS) is a devastating lesion of the renal allograft. De novo FSGS is uncommon compared to recurrent FSGS. AIM: To find out the incidence of de novo FSGS. MATERIALS AND METHODS: A retrospective evaluation of renal allograft biopsies was performed from 2007 to 2015, by light microscopy and immunohistochemistry including patient-donor demographics. Graft function status in terms of serum creatinine (SCr) and proteinuria were evaluated. RESULTS: Out of 2,599 renal allograft biopsies performed, 1.6% biopsies were reported as de novo FSGS. Majority were live related females donors with mean age of 43.8 years. Mean time of biopsy was 1.1 years post-transplant with proteinuria of 2.95 grams/24 hours and SCr of 2.24 mg/dL. Histopathological variants were collapsing 47.6%, Not Otherwise Specified/ classical 35.7%, cellular 9.5% and perihilar 7.1% biopsies. Associated Antibody Mediated Rejection (AMR) with T-Cell Rejection (TCR) was observed in 35.7% biopsies, acute on chronic CNI toxicity (calcineurin inhibitor) in five biopsies. Majority of the patients were on CNI based maintenance immunosuppression regimen. Total 28.6% patients and 23.8% grafts were lost over a mean follow up of 2.40 years. The mean SCr of remaining patients was 1.98 mg/dL. CONCLUSION: De novo FSGS can occur after the first year of renal transplant with related Human Leukocyte Antigen (HLA)matched donors leading to poor allograft survival. Close monitoring of urinary proteinuria and evaluation of allograft biopsy help in appropriate therapeutic modification to improve long term outcome of graft function.

Unusually Detected Anti-M Antibody Presenting as Cross Match Incompatibility in a Female Child Diagnosed with Small Round Cell Tumour.

MNS antigen system is one of the human blood group systems. Anti-M antibody is a relatively common, naturally occurring antibody of IgM variety. Clinically significant anti-M antibody is reactive at 37°C in the anti-human globulin phase due to high thermal amplitude of IgM component or presence of IgG component. If anti-M antibody is activated at 37°C or in the anti-human globulin phase, it may cause delayed haemolytic transfusion reactions or haemolytic disease of newborn, which suggest variable clinical significance. We report a case of an unusually detected anti-M antibody presenting as cross match incompatibility in a one-year-old female child with a lump in the right lumbar region, which was later diagnosed as small round cell tumour in the right kidney.

Stem cells: An emerging novel therapeutic for type-1 diabetes mellitus.

Stem cell based strategies are therapeutically potent for treating type-1 diabetes mellitus owing to their intrinsic regenerative capacity and immunomodulatory properties to arrest autoimmune ß-cell destruction, preserve residual ß-cell mass, facilitate endogenous regeneration, ameliorate innate/ alloimmune graft rejection, restore ß-cell-specific unresponsiveness in absence of chronic immunosuppression and to reverse hyperglycemia.

An unusual presentation of LCAT deficiency as nephrotic syndrome with normal serum HDL-C level.

Clinical and biochemical manifestations of lecithin-cholesterol acyltransferase (LCAT) deficiency include an abnormal lipid profile (characterized by hypercholesterolemia with markedly decreased high-density lipoprotein cholesterol [HDL-C] and hypertriglyceridemia), corneal opacities, hematologic abnormalities (normochromic anemia of varying intensity), splenomegaly, variable early coronary artery disease and nephropathy (initially proteinuria followed by progressive deterioration of renal function). We presented a patient with nephrotic syndrome, which renal biopsy revealed classic features of LCAT deficiency. To our knowledge, the present case is the first reported case of LCAT deficiency presenting with symptoms related to nephrotic syndrome in a patient with no obvious family history without any corneal deposits and normal HDL-C levels.

In-vitro generation of interleukin-10 secreting B-regulatory cells from donor adipose tissue derived mesenchymal stem cells and recipient peripheral blood mononuclear cells for potential cell therapy.

BACKGROUND: Interleukin-10 secreting B-cells are a major subset of B-regulatory cells (B-regs), commonly recognized as CD19+/38hi/24hi/IL10+. They carry out immunomodulation by release of specific cytokines and/or cell-to-cell contact. We have generated B-regs in-vitro from donor adipose tissue derived mesenchymal stem cells (AD-MSC) and renal allograft recipient (RAR) peripheral blood mononuclear cells (PBMC) for potential cell therapy. MATERIAL AND METHODS: Mononuclear cells separated by density gradient centrifugation from 50 ml anti-coagulated blood of 15-RAR and respective donors were analysed for baseline B-regs using appropriate antibodies. Equal amount (20 × 106 cells/ml) of stimulator (irradiated at 7.45 Gy/min for 10 min) and responder (non-irradiated) cells were co-cultured with in-vitro generated AD-MSC (1 × 106 cells/ml) in proliferation medium containing lipopolysaccharide from E. coli K12 strain at 37 °C with 5% CO2. Cells were harvested on day-7 and analyzed for viability, sterility, quantity, morphology and phenotyping. In-vitro generated B-reg levels were compared with baseline B-regs. RESULTS: In-vitro generated B-reg count increased to 16.75% from baseline count of 3.35%. CONCLUSION: B-regs can be successfully generated in-vitro from donor AD-MSC and RAR PBMC for potential cell therapy.