1. Huang lab resources to study COVID-19

A. Live SARS-CoV-2 infection systems for in vitro and in vivo studies:

Using the human lung epithelial cell line CALU-3, along with other more frequently used cells (VERO-E6, hACE2-A549, and hACE2-293T), we have developed a high-throughput flow-based assay for detecting SARS-CoV-2 infection. This system can be used to evaluate viral loads, neutralizing antibody titer, and antiviral drug efficacy. With the incorporation of a viability dye, our system also handily determine the cytopathic effects of the infection. An example of different variants of SARS-CoV-2 infection in CALU-3 cells are shown below:

B. Live SARS-CoV-2 infection in hACE2-KI mouse model for in vivo studies:

Along with the in vitro models above, we also use hACE2-KI mice to help better understand the effects of the vaccine and drug candidates of interest.

C. Pseudoviral system to study viral entry and screen for antiviral drugs:

We have established a pseudoviral platform to allow tests of spike-specific binding on viral particles, spike-mediated viral entry, neutralizing assays and anti-viral entry drug screening. The infection signal is about 10⁵ times over background, and the system is highly customizable for envelop and glyco- proteins of interest, including those found on SARS-CoV-2. Please contact us if this system is of interest to you.

♣ 1. Using this pseudoviral infection platform, we can perform neutralization assays to determine (1) the neutralizing potential of antibodies as therapeutic candidates; (2) whether people who recovered from COVIV-19 develop protective neutralizing antibodies; and (3) the potency of SARS-CoV-2 vaccine candidates in inducing protective neutralizing antibodies. An example of SARS-CoV-2 neutralizing antibody test is shown below:

♣ 2. This system does not involve live viruses, and is classified as biosafety level 2. We can customize the envelope protein to test highly transmissible and/or antibody-resistant viral strains and mutants. An example of SARS-CoV-2 spike D614G mutant is shown below:

2. Huang lab responses to COVID-19

January 12^th, 2020
A novel beta-coronavirus was isolated from SARS-like pneumonia patients in Wuhan China, and its genome sequence was first made accessible to the world by Dr. Fan Wu, Dr. Yongzhen Zhang in Shanghai Public Health Clinical Center and colleagues. This novel coronavirus emerging in 2019 was named 2019-nCoV.

January 20^th, 2020
Dr. Huang sent out an email with subject “SARS related”, to introduce SARS, 2019-nCoV outbreak, nCoV sequence published by Dr. Fan Wu, and phylogenetic clustering analyses.

January 30^th, 2020
Dr. Huang reached out to Dr. Gus Kousoulas and Dr. Paul Rider to discuss vaccine development strategy using VC2 vectors against 2019-nCoV.

January 31^st, 2020
Dr. Kousoulas held a nCoV-Pizza get together meeting to discuss the emerging coronavirus outbreak in China. Huang lab became part of the team in the arena.

February 11^th, 2020
The novel coronavirus was named by the International Committee on Taxonomy of Viruses as SARS-coronavirus 2 (SARS-CoV-2), and the associated disease was named by the World Health Organization as coronavirus disease (COVID-19). Dr. Huang met with department head Dr. Gus Kousoulas to discuss how to better advertise the terms “SARS-CoV-2” and “COVID-19”, and protective measures at work.

March 4^th, 2020
As one of the ten AAI public policy fellows, Dr. Huang visited the Capitol Hill, communicated with legislative representatives in Representative Garret Graves, Senator John Kennedy, and Senator Bill Cassidy’s offices. In these meetings, Dr. Huang advocated for NIH funding for biomedical research and supplemental bill support to help combat COVID-19.

March 16^th, 2020
The American Association of Immunologists had to cancel its annual meeting in Honolulu this May, due to COVID-19 pandemic. Huang lab has been preparing to attend the meeting with two selected oral presentations (by Dr. Natalie Nidetz and Mr. Michael McGee) and one block symposium co-chair (Dr. Weishan Huang), and is very sad about the situation. Cancellation is however a correct decision.
Huang lab uses mouse adapted H1N1 influenza viruses to study how flu infection can be blocked (inhibit viral entry), how immune responses to flu can be made specific (flu antigen-specific antibodies and T cells, induced by infections or vaccines), and how immunopathology caused by flu infection can be prevented and treated (molecular and cellular immunotherapies). We will continue these and start to investigate the similarities of viral pneumonia during flu and COVID-19.

March 23^rd, 2020
LSU moved to close campus following Gov. John Bel Edwards’ statewide ‘stay at home’ order. Huang lab wrapped up ongoing experiments, and turned all meetings virtual.

March 24^th, 2020
PhD student Michael McGee presented in lab meeting on the role of ITK in regulating IL-10 production in effector T cells and immunopathology during influenza A H1N1-induced viral pneumonia. The lab discussed strategies for effective work-from-home.

March 30^th, 2020
Huang lab learned as a group about COVID-19 Epidemiology, Pathophysiology, Diagnostics, Treatment, Prognosis, and Precautions.

April 6^th, 2020
Dr. Natalie Nidetz presented work on IL-10 function in innate immune regulation. And Dr. Huang presented retrospective research on COVID-19 diagnosis in China, to discuss the importance of integrating epidemiology, medical imaging, viral genomics, and immunology a comprehensive diagnosis.

April 13^th, 2020
Dr. Rezwanul Islam presented research on amino acid starvation signaling and TCR signaling on chronic allergic airway inflammation. Michael McGee reviewed current knowledge of immune profiling in COVID-19 patients. And Dr. Huang introduced the idea of using computational analyses to better understand susceptibility to SARS-CoV-2 of different animal species. We also discussed the potential of Huang lab in COVID-19 research to help contribute to a better understanding and control of the virus and disease.

April 20^th, 2020
Dr. Huang presented the role of non-receptor tyrosine kinase (NRTK) ITK signaling in Foxp3+ Treg cell stability and function, while Michael McGee discussed the potential role of ITK and other NRTK signaling in viral pneumonia associated with COVID-19.

April 25^th, 2020
Dr. Huang published her article “COVID-19 diagnostic process in mainland China: the math beyond pneumonia” in Journal of Allergy and Clinical Immunology.

April 26^th, 2020
Michael McGee presented in lab meeting journal club to discuss the role of eosinophils and allergic airway inflammation in influenza A H1N1 growth and the flu-induced lung immunopathology.

May 4^th, 2020
In lab meeting today, Dr. Huang discussed the therapeutic effects of inhibiting ITK signaling in allo-bone marrow transplantation, to retain graft-versus-tumor effect but prevent graft-versus-host disease. Dr. Huang also presented an article to help the lab learn about designer DNA for Pattern-recognition Enabled Sensing and Therapeutics (PEST), which may help develop strategies for diagnosis, blockade and vaccination against infections. Dr. Rezwanul Islam presented a journal club article to discuss signaling pathways that regulate inflammatory ILC2-17 development and function in the airway. Michael McGee presented Dr. Jie Sun’s research article on the role of CD4⁺ T cells and innate immune cell-derived IL-27 in IL-10 production by flu antigen-specific CTLs.

May 11^th, 2020
Dr. Natalie Nidetz presented her research on GCN2 signaling in mast cell and macrophage-mediated inflammation. The lab also discussed how CRISPR-based gene detection methods can be utilized to help combat COVID-19.

May 18^th, 2020
Huang lab is approved to return to essential research on campus at LSU. Dr. Huang presented in lab meeting a collaborative research with doctors in Wuhan and Guangzhou to investigate the kinetics and level of SARS-CoV-2-specific IgG in COVID-19 patients.