Endometrial Staining of CD56 (Uterine Natural Killer), BCL-6, and CD138 (Plasma Cells) Improve Diagnosis and Clinical Pregnancy Outcomes in Unexplained Infertility and Recurrent IVF Failures: Standardization of Diagnosis with Digital Pathology

Abstract

In women with unexplained infertility (UI) and recurrent in vitro fertilization (IVF) failures, the etiology is often unclear. Endometrial immune perturbations and the use of immune markers associated with these dysregulations are of great interest in the diagnosis and treatment of UI. However, reliable biomarkers and standardized quantification methods are lacking. Here, to address endometrial immune dysregulation in UI patients with recurrent IVF failures, we performed endometrial tissue sampling and immunostaining of CD56 (uNK), CD138, and BCL-6. Of these cases, 57.9% had positive CD56 in the endometrial stroma, while 46.1% had positive BCL-6 in the glandular epithelium, and 14.5% of the cases were found to be positive for CD138. Combined staining rates were 60.5%, 68.4%, and 71.05% for (CD56 or BCL-6), (CD56 or CD138), and (CD56, BCL-6, or CD138), respectively. There was a significant correlation between CD56 and BCL-6 positivity, while CD138 positivity was an independent parameter. After the recommended targeted therapy, pregnancy rates were found to increase from 58.5% to 61.6% and 73.8% in CD56-positive, (CD56- or BCL-6-positive), and (CD56-, BCL-6-, or CD138-positive) cases, respectively. Notably, a retrospective evaluation of digital pathology and light microscopy results showed a significant correlation. This study suggests that the examination of CD56, BCL-6, and CD138 in the same endometrial sample may be an effective method in determining the etiology of UI and reaching an early diagnosis and treatment options. Moreover, digital pathology can be used in the evaluation of CD56 and BCL-6 to provide objective, rapid, and reliable results.

The Programmed Cell Death Ligand 1 and Lipocalin 2 Expressions in Primary Breast Cancer and Their Associations with Molecular Subtypes and Prognostic Factors

“…Three micrometers thick sections were prepared from the blocks and stained with hematoxylin and eosin (H&E) staining using a Shandon Gemini stainer as described previously. 30 Immunohistochemical staining was performed in accordance with standard and validated immunohistochemical protocols. Briefly, following antibodies were used: antibodies against ER (ER-6F11-L-CE, Clone 6f11, Leica Biosystems), PR (PGR-312/2-L-CE Clone 16, Leica Biosystems), HER-2/neu4B5 (anti-HER-2/neu (4B5), Rabbit Monoclonal Primary Antibody, Ventana, Roche), and Ki-67 (Monoclonal Mouse anti-Human Clone MIB-1, Dako) using a Ventana Benchmark XT device (Roche Diagnostics, Basel, Switzerland).…”

“…Briefly, following antibodies were used: antibodies against ER (ER-6F11-L-CE, Clone 6f11, Leica Biosystems), PR (PGR-312/2-L-CE Clone 16, Leica Biosystems), HER-2/neu4B5 (anti-HER-2/neu (4B5), Rabbit Monoclonal Primary Antibody, Ventana, Roche), and Ki-67 (Monoclonal Mouse anti-Human Clone MIB-1, Dako) using a Ventana Benchmark XT device (Roche Diagnostics, Basel, Switzerland). 30 …”

“…The PD-L1 (Clone: CAL10, Dilution: 1/100 Brand: Biocare Medical) and LCN2 (Polyclonal, Dilution: 1/200 Brand: GeneTex) were immunohistochemically applied to the same tumor blocks with the Ventana Benchmark XT device (Roche Diagnostics, Basel, Switzerland). 30 Negative and positive controls were used for both immune markers. All pictures were taken by Viracenter Digital Pathology new version.…”

“…Briefly, following antibodies were used: antibodies against ER (ER-6F11-L-CE, Clone 6f11, Leica Biosystems), PR (PGR-312/2-L-CE Clone 16, Leica Biosystems), HER-2/neu4B5 (anti-HER-2/neu (4B5), Rabbit Monoclonal Primary Antibody, Ventana, Roche), and Ki-67 (Monoclonal Mouse anti-Human Clone MIB-1, Dako) using a Ventana Benchmark XT device (Roche Diagnostics, Basel, Switzerland). 30 The Nottingham modification of Bloom Richardson Grading was used for tumor histological and nuclear grading. 31 In addition, prognostic factors such as tumor diameter, lymphovascular invasion, necrosis, and lymph node metastasis were noted for each case.…”