Formalin fixation for optimal concordance of programmed death-ligand 1 immunostaining between cytologic and histologic specimens from patients with non-small cell lung cancer.

BACKGROUND: Immunohistochemical staining of programmed death-ligand 1 (PD-L1) is used to determine which patients with non-small cell lung cancer (NSCLC) may benefit most from immunotherapy. Therapeutic management of many patients with NSCLC is based on cytology instead of histology. In this study, concordance of PD-L1 immunostaining between cytology cell blocks and their histologic counterparts was analyzed. Furthermore, the effect of various fixatives and fixation times on PD-L1 immunoreactivity was studied. METHODS: Paired histologic and cytologic samples from 67 patients with NSCLC were collected by performing fine-needle aspiration on pneumonectomy/lobectomy specimens. Formalin-fixed, agar-based or CytoLyt/PreservCyt-fixed Cellient cell blocks were prepared. Sections from cell blocks and tissue blocks were stained with SP263 (standardized assay) and 22C3 (laboratory-developed test) antibodies. PD-L1 scores were compared between histology and cytology. In addition, immunostaining was compared between PD-L1-expressing human cell lines fixed in various fixatives at increasing increments in fixation duration. RESULTS: Agar cell blocks and tissue blocks showed substantial agreement (κ = 0.70 and κ = 0.67, respectively), whereas fair-to-moderate agreement was found between Cellient cell blocks and histology (κ = 0.28 and κ = 0.49, respectively). Cell lines fixed in various alcohol-based fixatives showed less PD-L1 immunoreactivity compared with those fixed in formalin. In contrast to SP263, additional formalin fixation after alcohol fixation resulted in preserved staining intensity using the 22C3 laboratory-developed test and the 22C3 pharmDx assay. CONCLUSIONS: Performing PD-L1 staining on cytologic specimens fixed in alcohol-based fixatives could result in false-negative immunostaining results, whereas fixation in formalin leads to higher and more histology-concordant PD-L1 immunostaining. The deleterious effect of alcohol fixation could be reversed to some degree by postfixation in formalin.

Hypercoiling of the umbilical cord and placental maturation defect: associated pathology?

Our objective was to determine whether there is an association between hypercoiling of the umbilical cord and placental maturation defect. From a database comprising 1147 cases, containing data on all placentas examined at our institution during the study period, we selected all cases with a gestational age of at least 37 weeks that exhibited hypercoiling of the umbilical cord (coiling density above the 90th percentile, n = 42); we also examined 2 matched controls for each case, one with hypocoiling and one with normocoiling. The mean number of syncytiocapillary membranes (SCM) per terminal villus was calculated. Presence of a placental maturation defect was defined as the mean number of SCM below the 10th percentile. Correlations were assessed using Spearman's rho. Relations between dichotomous variables were tested using logistic regression. Mean number of SCM per terminal villus (+/-standard deviation) was 1.25 +/- 0.65. Difference in mean between hypo- and hypercoiled cords was 0.37 (95% confidence interval [CI], 0.07 to 0.67). The correlation coefficient between mean number of SCM and umbilical coiling index (UCI, coils/cm) was -0.28 (P = 0.002). The odds ratio (OR) for placental maturation defect in presence of hypercoiling was 2.61 (95% CI, 0.75 to 9.12). The OR for fetal death was 132 (95% CI, 13.2 to 1315) in the presence of a placental maturation defect and 5.49 (95% CI, 1.02 to 29.6) in the presence of hypercoiling. The OR for indication of fetal hypoxia/ischemia was 12.3 (95% CI, 3.0 to 50.3) in the presence of a placental maturation defect and 3.2 (95% CI, 0.95 to 10.9) in the presence of hypercoiling. We found a trend toward placental maturation defect in the presence of hypercoiling and an inverse relationship between the mean number of SCM in the terminal villi and the UCI. We confirmed associations between fetal death and both a maturation defect and hypercoiling and found an association between histological indication of fetal hypoxia/ischemia and a placental maturation defect.