A Method for in Vivo Quantification Of Cytokine IL-1β In The Rat Intrathecal Space

Abstract

IL-1β is a potent pro-inflammatory cytokine critical to multiple pathologies in the central nervous system (CNS). Quantification of IL-1β in vivo is challenging due to pM range of IL-1β released in the spinal cord and also the terminal nature of cerebrospinal fluid (CSF) sampling in rodents. Herein we developed a robust in vivo device on stainless steel suitable for detection of IL-1β in the spinal cord of rats. This approach offers high sensitivity (3.2 pg mL −1 ) and specificity to IL-1β. Also, a modified lumbar puncture method was employed to implant the device in the intrathecal space of male Sprague−Dawley (SD) rats under short-acting anesthesia, allowing minimal invasiveness, which provided the possibility of repeated measurement of IL-1β in the same animal. Our biosensing technology and the surgical method provide a universal platform for in vivo detection of diverse analytes in longitudinal, within-subject studies in the intrathecal space of rats to reduce the required number of experimental animals.

Cytokines: From Clinical Significance to Quantification

“…[31,126] They can also use other transducing elements. TNF-𝛼 FI 20 pg mL −1 10 5 -10 6 pg mL −1 -Near real time [227] IFN-𝛾 FI 1.5 × 10 4 pg mL −1 (0.2-8) × 10 5 pg mL −1 -≈6 h [130] IFN-𝛾 FI 2 pg mL −1 5-10 2 pg mL −1 -≈30 min [33] IFN-𝛾 FI 0.1 pg mL −1 0.1-1.5 × 10 3 pg mL −1 -≈2 h [131] IFN-𝛾 FI 2 pg mL −1 0-10 2 pg mL −1 -≈45 min [228] IFN-𝛾, TNF-𝛼 FI 21 pg mL −1 0-3.6 × 10 2 pg mL −1 -≈40 min [128] IL-1𝛽 FI 3.2 pg mL −1 3.5-2 × 10 2 pg mL −1 5-10 µL - [35] IL-20 FI 0.2 pg mL −1 2-2 × 10 4 pg mL −1 5 µL - [229] IL-1𝛽 FI 4.7 pg mL −1 13-2 × 10 2 pg mL −1 1 µL - [230] IL-1𝛽 FI 10 pg mL −1 25-4 × 10 2 pg mL −1 -- [125] IL-6 FI 1 pg mL −1 1-4 × 10 2 pg mL −1 1 µL - [37] IL-6 FI 0.1 pg mL −1 0.4-4 × 10 2 pg mL −1 1 µL - [231] IFN-𝛾 FI 10 3 pg mL −1 5 × 10 3 -1 × 10 5 pg mL −1 -Near real time [32] IL-2, IL-4, IL-6 SPR 5-20 pg mL −1 10-10 4 pg mL −1 1 µL ≈40 min [24] IL-6 SPR 10 pg mL −1 10-10 2 pg mL −1 -≈30 min [232] IL-6, TNF-𝛼 SPR 5 pg mL −1 4-5 × 10 2 pg mL −1 -- [137] IL-6, IL-4, IL-10,TNF-𝛼 SPR 20 pg mL −1 10-10 4 pg mL −1 1 µL ≈30 min [233] IFN-𝛾 SPR 5 × 10 4 pg mL −1 (0.5-8) × 10 5 pg mL −1 800 µL Real time [136] IL-6 SPR 10 4 pg mL −1 10 4 -2 × 10 5 pg mL −1 100 µL Real time [138] TGF-𝛽1 E C 1 0 p g m L −1 15-3 × 10 3 pg mL −1 25 µL ≈60 min [234] IL-6, IL-1𝛽, TNF-𝛼 EC 5 pg mL −1 5-2 × 10 2 pg mL −1 -- [28] IFN-𝛾 EC 1.6 pg mL −1 2.5-2 × 10 3 pg mL −1 5 µL ≈200 s [152] IFN-𝛾 EC 0.2 ng mL −1 0.2-2.8 × 10 2 ng mL −1 -- [156] IFN-𝛾 EC 3 pg mL −1 10-5 × 10 3 pg mL −1 30 µL ≈60 min [155] TNF-𝛼 EC -1-15 pg mL −1 -- [235] IFN-𝛾 EC 6 pg mL −1 10-5 × 10 2 pg mL −1 100 µL Real time [31] VEGF EC 0.1 pg mL −1 2-5 × 10 2 pg mL −1 -Real time [126] TNF-𝛼 EC 0.1 pg mL −1 0.1-1.5 × 10 2 pg mL −1 -≈20 min [29] IL-1𝛽, IL-10 EC 0.3 pg mL −1 (IL-10) 0.7 pg mL −1 (IL-1𝛽)…”

“…Aptamers have also garnered interest in biosensing applications due to their small size, reusability as compared to single use antibodies and efficient immobilization at high density. [26] A variety of biosensing platforms for quantification of cytokines ranging from sandwich immunosensors [28][29][30] to aptasensors, [31,32] nanosensors [33,34] implantable medical devices, [30,[35][36][37] point-ofcare (POC) diagnostics, [31] in vivo real-time monitoring, [38] and from intracellular bioimaging [39] to extracellular detection [40] have been reported. This review will introduce cytokines from the perspective of the pathways they trigger and whether they are inflammatory or anti-inflammatory.…”

Universal CRISPR/Cas12a-associated aptasensor suitable for rapid detection of small proteins with a plate reader

“…It is simple, universal, versatile, and compatible with current ELISA measurement setups, such as plate readers. Moreover, as aptamers are generally not temperature-sensitive and can work at various temperature ranges, such as 37 °C (Munzar et al, 2019), it shows potential to be adapted for in vivo diagnostic applications, e.g., using an implantable device for IL-1β detection, as reported by Deng et al (2020) Meanwhile, as both the aptamer and Cas12a proteins retain their functions on paper-based substrates, the CAMERA system shows potential to be transferred to microfluidic paper-based analytical devices, such as paper lateral flow assay test strips, to realize the POC diagnostics. Although applying the aptamer may raise potential limitations on its availability to different analytes and sensitivity to nuclease contaminations, which can be relieved with the further expanded aptamer pool and additional nucleic acid modification techniques, we expect that the CAMERA method will inspire the use of CRISPR/Cas biosensing technology in combining with other existing biosensing components for novel biosensor development.…”

Point‐of‐care detection of cytokines in cytokine storm management and beyond: Significance and challenges

“…have developed different biosensing platforms for single cytokine monitoring from in vitro to in vivo with fluorescence signal readout [ 78–84 ] or electrochemical signal readout. [ 85,86 ] In order to realize the cytokine monitoring in mouse brain or spinal cords, deployable devices based on immunosensors on optical fiber [ 87,88 ] and stainless steel [ 80,89 ] have been developed for detection of spatially localized cytokines at the levels of pg/mL. An impedance aptasensor was developed for highly sensitive and selective detection of IL‐6 with a good linear response from 5 pg/mL to 100 ng/mL and a detection limit of 1.6 pg/mL.…”