Pre-partum blood leukocyte profile distinguishes gestational inflammatory stages to prognosticate birth-related adverse outcomes

Hari Shankar^1,2*Yash Gupta^3*Neeta Kumar¹Vinoth Gnana Chellaiyan Devanbu^4*William M Trochim⁵Almira Lydia Hoogesteijn⁶Jeanne Marie Fair⁷Raja Babu Singh Kushwah⁸Michelle Iandiorio⁹Andrew Yen⁵Donthamsetty Nageswara Rao^2*Ariel L Rivas^9*

• ¹Indian Council of Medical Research (ICMR), New Delhi, India
• ²All India Institute of Medical Sciences New Delhi, New Delhi, India
• ³Penn State College of Medicine, Hershey, United States
• ⁴Chettinad Hospital and Research Institute, Kanchipuram, India
• ⁵Cornell University, Ithaca, United States
• ⁶Cinvestav Unidad Merida, Mérida, Mexico
• ⁷Los Alamos National Laboratory, Los Alamos, United States
• ⁸Texas A&M University, College Station, United States
• ⁹The University of New Mexico, Albuquerque, United States

Introduction: Pregnancy is a physiological process accompanied by immuno-dynamic changes (inflammatory stages) that could influence or predict pregnancy outcomes. Yet, ambiguity (overlapping data intervals on biomedically different conditions) may prevent such differentiation. Here, a retrospective, proof-of-concept study was conducted to (a) differentiate pregnancy-related inflammatory stages with ambiguity, and (b) to prognosticate birth-related double risks (low birth weight and pre-term birth) based on blood tests of pregnant women. Method – Blood samples collected from 131 Indian pregnant females (192 temporal observations) were retrospectively analyzed with: (1) a reductionist approach, which investigates cell types individually; and (2) a non-reductionist alternative, which uses a proprietary software package to explore pre-partum multicellular interactions and birth-related outcomes. Leukocyte percentages collected during the second and third trimesters was utilized to predict double risks. Results – While the reductionist analysis failed to distinguish double risks (ambiguity was observed), the non-reductionist method differentiated four inflammatory stages, characterized by: (i) no double risk and a high phagocyte/lymphocyte (P/L) ratio (class ‘A’), (ii) no double risk and a very low P/L ratio (class ‘B’), (iii) 16.6% double risks and a moderately elevated P/L ratio (class ‘C’), and (iv) 83.3% double risks and the highest monocyte percentage (class ‘D’). All double risks observations were associated with statistically higher concentrations of serum ferritin. Discussion – Combined, longitudinal clinical-inflammatory and personalized data patterns inform whether a pregnancy is associated with double risks and/or when changes occur. Because pre-partum observations anticipated birth-related outcomes, personalized and prognostic features were demonstrated. Since antenatal care involves routine blood sampling (a low-cost procedure), this methodology is inherently translational. Because construct, internal, external, and statistical validity were supported, if corroborated with prospective studies, this method may assist United Nations’ 2023 goals toward reducing infant mortality.