Strong Immunity Research Articles

A Study on Effectiveness of Yoga for Improvement of Work Productivity among Millennials in Tiruchirappalli

Yoga has been great art and science of healthy living. Positive health is an important means of leading meaningful life filled with happiness and inner harmony. With rapid economic development and increasing westernization of lifestyle in past few decades there is a change in the behavioural pattern of human being. This article focuses on the effectiveness of yoga for improvement of work productivity among millennials. The regular practice of yoga is a solution for the many psychological and lifestyle disorders of the modern life with the revitalization of the traditional inheritance along the lines of the sustainable approach. It was observed that, all the responders said that yoga is beneficial in their life as it helps them to attain inner peace, productive mind, strong immunity and balanced work life as well. Thus, it can be concluded that, practicing yoga makes life healthy and increased their productivity.

Intranasal immunization of mice with chimera of Salmonella Typhi protein elicits protective intestinal immunity

Development of safe, highly effective and affordable enteric fever vaccines is a global health priority. Live, oral typhoid vaccines induce strong mucosal immunity and long-term protection, but safety remains a concern. In contrast, efficacy wears off rapidly for injectable, polysaccharide-based vaccines, which elicit poor mucosal response. We previously reported Salmonella Typhi outer membrane protein, T2544 as a potential candidate for bivalent (S. Typhi and S. Paratyphi A) vaccine development. Here, we show that intranasal immunization with a subunit vaccine (chimera of T2544 and cholera toxin B subunit) induced strong systemic and intestinal mucosal immunity and protection from S. Typhi challenge in a mouse model. CTB-T2544 augmented gut-homing receptor expression on lymphocytes that produced Th1 and Th17 cytokines, secretory IgA in stool that inhibited bacterial motility and epithelial attachment, antibody recall response and affinity maturation with increased number of follicular helper T cells and CD4+ central and effector memory cells.

Is Post-COVID 19 Vaccination Antibody Level Related to Happiness and Stress Hormones?

Aim: The level of antibody production in those vaccinated against coronavirus can be affected by many different situations. There is an important balance between immune response, stress and emotional state. However, it is not known how this situation affects antibody production after vaccination. This study aimed to investigate the correlation between the antibody response induced by the COVID-19 vaccine and the hormones cortisol, a marker of the stress axis, and serotonin, a marker of happiness. 
 Material and Method: Serum cortisol and serotonin levels were analyzed in those who tested positive (n=40) and negative (n=40) for Anti-SARS CoV-2 IgG induced by vaccination. Anti-SARS CoV 2 IgG, cortisol, and serotonin levels were determined by using the ELISA method. The data were compared using the Mann-Whitney U test. The value of p0.05). 
 Conclusion: Consequently, no effect of the stress parameter cortisol and the happiness parameter serotonin, was found in vaccine-induced immunization. It is considered that the different antibody responses in individuals may vary depending on other factors.

Immune responses to mRNA-based vaccines given as athird COVID-19 vaccine dose in people living with HIV-a literature review.

People living with HIV (PLWH) were not included in the first efficacy studies of mRNA vaccines against SARS-CoV-2. In this literature review, we investigate evidence of humoral and cellular immunity after a third dose of an mRNA vaccine in PLWH. We performed a literature search in PubMed, Embase, Web of Science and SCOPUS published between 1 January 2020 and 31 December 2022. Selection criteria were studies on immunological responses in PLWH, who were given an mRNA-based vaccine as a third vaccine dose against SARS-CoV-2. Eight articles complied with our selection criteria. All studies found a strong humoral response after the third dose. Five studies investigated cellular immunity and found an increased cellular response after the third vaccine dose in PLWH. No difference in humoral response was observed between PLWH and controls after three doses. However, some of the studies suggested a weaker cellular response among PLWH than in controls, which was associated with lower nadir or current CD4+ T-cell counts. In conclusion, we found evidence of strong humoral immunity in PLWH after receiving an mRNA-based COVID-19 vaccine as a third dose, while the cellular immunity may be impaired compared to controls.

A rabies mRNA vaccine with H270P mutation in its glycoprotein induces strong cellular and humoral immunity

Rabies is a lethal zoonotic disease that kills approximately 60,000 people each year. As the sole virion-surface protein, the rabies virus glycoprotein (RABV-G) mediates its host-cell entry. RABV-G’s pre-fusion conformation displays major known neutralizing antibody epitopes, which can be used as immunogen for prophylaxis. H270P targeted mutation can stabilize RABV-G in the pre-fusion conformation. Herein, we report the development of a highly promising rabies mRNA vaccine composed of H270P targeted mutation packaged in lipid nanoparticle (LNP), named LNP-mRNA-G-H270P. Humoral and cellular immunity of this vaccine were assessed in mice comparing to the unmodified LNP-mRNA-G and a commercially available inactivated vaccine using one-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparisons test. The results show the titer of RABV-G-specific IgG and virus-neutralization antibody titers (VNTs) in LNP-mRNA-G-H270P group were significant higher than those in LNP-mRNA-G and inactivated vaccine groups. Likewise, IFN-γ-secreting splenocytes, level of IL-2 in the supernatant of spleen cells, as well as IFN-γ-producing CD4+ T cells in LNP-mRNA-G-H270P group were significant higher than those in the other two vaccine groups. Hence, these results demonstrated that targeting the H270P mutation in RABV-G through an mRNA-LNP vaccine platform represents a promising strategy for developing a more efficacious rabies vaccine.

Rolling bearing fault diagnosis based on variational mode decomposition and weighted multidimensional feature entropy fusion

Since bearing fault signal in complex running status is usually characterized as nonlinear and non-stationary, it is difficult to extract accurate affluent features and achieve effective fault identification via conventional signal processing tools. In this article, a rolling bearing fault diagnosis technique based on variational mode decomposition and weighted multidimensional feature entropy fusion is proposed to address this issue, which is mainly composed of three procedures. First, the original signal undergoes the variational model decomposition. Next, the signal features are extracted by weighted multidimensional feature entropy as the input of the diagnosis model. Finally, the classification is performed by a convolutional neural network. The method is applied in simulation and experimental analysis. The experimental results show that the proposed method, which demonstrates strong immunity to noise and robustness, can more effectively and adaptively extract the fault features of rolling bearings and achieve the goal of identifying the rolling bearing fault category and damage degree under variable operating conditions. Meanwhile, this approach exhibits superior accuracy and identification performance to some similar entropy-based hybrid approaches referred to in this article, with a promising prospect in industrial application.

Multi-signal sensor array based on a fluorescent nanozyme for broad-spectrum screening of pesticides

Accurate identification of broad-spectrum pesticides remains a challenge for existing rapid detection technologies based on natural cholinesterase. Sensor arrays show excellent promise, but typically rely on increasing receptors to improve sensing dimensionality, which is both cumbersome and imprecise. In this work, inspired by the multiple properties of a fluorescent nanozyme, we constructed a facile and simple sensor array based on the laccase-like activity, peroxidase-like activity, and fluorescent feature of one nanozyme (Cu-BDC-NH2). In the presence of pesticides, they produced differential positive responses and could be captured by linear discriminant analysis and hierarchical clustering analysis. Thus, different pesticides belonged to organophosphates, carbamates, organochlorines, pyrethroids, sulfonylureas, and diphenyloxazolines, respectively, have been successfully discriminated at 1 – 100 μg/mL. The differentiation of cross-contamination at different concentrations and compositions further validated the sensing performance. More importantly, they demonstrated strong immunity to interference as well as high feasibility for recognizing pesticides in fruits and vegetables, and performed precise identification for blind samples. Therefore, the sensor arrays provided a novel and reliable solution for broad-spectrum pesticide screening, which not only bridged the gap of existing rapid detection methods, but also opened up a new way for food safety regulation.

Cancer cell membrane proteins-encapsulated nanovaccine enhances cancer immunotherapy and prevention by provoking antigen-specific cellular immunity via the dendritic cell-targeted delivery

Cancer nanovaccines offer a promising strategy for fighting against tumors, however, the engineering of cancer nanovaccines that can be easily fabricated with tanglesome cancer cell-derived antigens and elicit an adequately strong tumor-specific cellular immunity remains challenging. Herein, metal-phenolic networks (MPNs) are used as an antigen delivery platform to prepare the mannose-modified MPNs nanovaccine loaded with ovalbumin (OVA) and the immunoadjuvant CpG oligodeoxynucleotide (MMOC) through the facile self-assembly. When the model antigen OVA is substituted with cancer cell membrane proteins (CCMPs), the nanovaccine is called MMCC. MMOC markedly activates dendritic cells (DCs) via the mannose-mediated endocytosis and efficiently promotes the antigen cross-presentation, thus inspiring a robust antigen-specific CD8+ T cell response as well as immune memory effect in vivo. Consequently, MMOC exhibits admirable therapeutic and preventive results on E.G7-OVA tumors. Moreover, the combination use of MMCC with anti-PD1 significantly inhibits the growth of 4T1 tumors by strengthening the cellular immunity and decreasing the proportion of regulatory T cells (Tregs). The survival rate of 4T1 tumor-bearing mice in the prophylaxis assay is maintained at 100 % by MMCC over 42 days. Altogether, this study affords a universal and effective nanovaccine preparation strategy for cancer immunotherapy and prevention.

A Self-Adaptive Pyroptosis Inducer: Optimizing the Catalytic Microenvironment of Nanozymes by Membrane-Adhered Microbe Enables Potent Cancer Immunotherapy.

Pyroptosis has garnered increasing attention in cancer immunotherapy. Moreover, increasing plasma membrane damage by reactive oxygen species (ROS) is considered an effective strategy for promoting pyroptosis. However, the current tactics for enhancing membrane rupture in pyroptosis are limited by the inherent drawbacks of ROS and the immunosuppressive tumor microenvironment. Herein, a self-adaptive pyroptosis inducer (LPZ) is designed by integrating Lactobacillus rhamnosus GG (LGG) and an enzyme-like metal-organic framework to achieve potent pyroptosis immunotherapy. LPZ can adhere to cancer cell membranes through the interaction between the pili of LGG and the mucin of cancer cells. In particular, the adaptive formula can gradually enhance the ability of nanozymes to produce ROS by creating an acidic microenvironment through anaerobic respiration. These results verify that LPZ could generate high levels of ROS both on the membrane and within cancer cells, leading to pyroptotic cell death and strong antitumor immunity. Meanwhile, LGG are eventually killed by ROS in this process to halt their respiration and prevent potential biosafety concerns. Overall, this work provides new inspiration for the design of self-adaptive nanocatalytic drugs for cancer immunotherapy.

A potential dual protection vaccine: Recombinant feline herpesvirus-1 expressing feline parvovirus VP2 antigen

Recently, herpesvirus viral vectors that stimulate strong humoral and cellular immunity have been demonstrated to be the most promising platforms for the development of multivalent vaccines, because they contain various nonessential genes and exhibit long-life latency characteristics. Previously, we showed that the feline herpesvirus-1 (FHV-1) mutant WH2020-ΔTK/gI/gE, which was safe for felines and provided efficacious protection against FHV-1 challenge, can be used as a vaccine vector. Moreover, previous studies have shown that the major neutralizing epitope VP2 protein of feline parvovirus (FPV) can elicit high levels of neutralizing antibodies. Therefore, to develop a bivalent vaccine against FPV and FHV-1, we first generated a novel recombinant virus by CRISPR/Cas9-mediated homologous recombination, WH2020-ΔTK/gI/gE-VP2, which expresses the VP2 protein of FPV. The growth characteristics of WH2020-ΔTK/gI/gE-VP2 were similar to those of WH2020-ΔTK/gI/gE, and WH2020-ΔTK/gI/gE-VP2 was stable for at least 30 generations in CRFK cells. As expected, we found that the felines immunized with WH2020-ΔTK/gI/gE-VP2 produced FPV-neutralizing antibody titers (27.5) above the positive cutoff (26) on day 14 after single inoculation. More importantly, recombinant WH2020-ΔTK/gI/gE-VP2 exhibited severely impaired pathogenicity in inoculated and cohabiting cats. The kittens immunized with WH2020-ΔTK/gI/gE and WH2020-ΔTK/gI/gE-VP2 produced similar levels of FHV-specific antibodies and IFN-β. Furthermore, felines immunized with WH2020-ΔTK/gI/gE-VP2 were protected against challenge with FPV and FHV-1. These data showed that WH2020-ΔTK/gI/gE-VP2 appears to be a potentially safe, effective, and economical bivalent vaccine against FPV and FHV-1 and that WH2020-ΔTK/gI/gE can be used as a viral vector to develop feline multivalent vaccines.

Effect of milking time on yield, composition, and antimicrobial components of milk in lactating goats.

Various studies have attempted to improve the milk yield and composition in dairy animals. However, no study has examined the effects of milking at different times on milk yield and composition. Therefore, this study aimed to compare the yield, composition, and antimicrobial components of milk obtained from milking at different times in lactating goats. Eight goats were milked once daily at different times for three consecutive weeks (first week: 06:00 h; second week: 09:00 h; and third week: 12:00 h). The light ranged from 06:30 to 19:00 h. Milk and blood samples were collected once a day during milking time. Milking at 09:00 h resulted in a significantly higher milk yield than that obtained after milking at 06:00 and 12:00 h. Prolactin levels in plasma and the fat, Na+, β-defensin, and S100A7 (antimicrobial component) levels in milk were the lowest in the 09:00 h milking. These results indicate that milk yield, composition, and antimicrobial components can be affected by milking time, which may be related to the altered concentration of prolactin in the blood. These findings provide a rational basis for achieving maximal milk production with strong immunity by changing to a more effective milking time.

Th2 and Th17-associated immunopathology following SARS-CoV-2 breakthrough infection in Spike-vaccinated ACE2-humanized mice.

Vaccines have demonstrated remarkable effectiveness in protecting against COVID-19; however, concerns regarding vaccine-associated enhanced respiratory diseases (VAERD) following breakthrough infections have emerged. Spike protein subunit vaccines for SARS-CoV-2 induce VAERD in hamsters, where aluminum adjuvants promote a Th2-biased immune response, leading to increased type 2 pulmonary inflammation in animals with breakthrough infections. To gain a deeper understanding of the potential risks and the underlying mechanisms of VAERD, we immunized ACE2-humanized mice with SARS-CoV-2 Spike protein adjuvanted with aluminum and CpG-ODN. Subsequently, we exposed them to increasing doses of SARS-CoV-2 to establish a breakthrough infection. The vaccine elicited robust neutralizing antibody responses, reduced viral titers, and enhanced host survival. However, following a breakthrough infection, vaccinated animals exhibited severe pulmonary immunopathology, characterized by a significant perivascular infiltration of eosinophils and CD4+ T cells, along with increased expression of Th2/Th17 cytokines. Intracellular flow cytometric analysis revealed a systemic Th17 inflammatory response, particularly pronounced in the lungs. Our data demonstrate that aluminum/CpG adjuvants induce strong antibody and Th1-associated immunity against COVID-19 but also prime a robust Th2/Th17 inflammatory response, which may contribute to the rapid onset of T cell-mediated pulmonary immunopathology following a breakthrough infection. These findings underscore the necessity for further research to unravel the complexities of VAERD in COVID-19 and to enhance vaccine formulations for broad protection and maximum safety.

Gold standard assessment of immunogenic cell death induced by photodynamic therapy: From in vitro to tumor mouse models and anti-cancer vaccination strategies.

The discovery of the concept of immunogenic cell death (ICD) is a cornerstone in the development of novel anti-cancer immunotherapeutic approaches. Induction of the ICD pathway by specific anti-cancer therapeutic regimens can eliminate cancer cells by directly killing them during therapy and by activation of strong and specific anti-cancer immunity, leading to a long-lasting immunological memory that prevents cancer recurrence. ICD encompasses different forms of regulated cell death and can be triggered by many anti-cancer treatment modalities, including photodynamic therapy (PDT). PDT is a multistep procedure involving the accumulation of a light-sensitive dye known as a photosensitizer (PS) in tumor cells, followed by its activation by irradiation with a light of an appropriate wavelength. In the presence of molecular oxygen, the irradiated PS leads to the generation of cytotoxic reactive oxygen species, which can lead to ICD induction in the cancer cells. Here, we first describe in vitro methods to help optimize the PDT procedure for a specific PS. We also provide a collection of protocols and techniques for assessing ICD in vitro, including analysis of the emission of damage associated molecular patterns (DAMPs), efferocytosis, and the maturation and activation state of antigen presenting cells. Next, we describe in detail protocols for diverse tumor mouse models for assessing and characterizing ICD in vivo, such as murine tumor vaccination models. Finally, as an immunotherapeutic vaccine, we suggest using either PDT-induced dead cancer cells, preferably undergoing ICD, or dendritic cells loaded with lysates of PDT-induced cancer cells in a syngeneic orthotopic glioma model. Overall, this methodological article provides a quantitative, comprehensive set of validated tools that can be successfully used, with some adaptations, to identify, optimize and validate novel PSs in vitro and in vivo for the efficient induction of ICD during photodynamic treatment.

Circadian Genes and Economic Traits in Livestock Animals

Circadian rhythms are oscillators of endogenous autonomic activity in all living organisms and regulate economic traits such as reproduction, milk characteristics and growth performance in farm animals. These rhythms are directly or indirectly controlled by the circadian clock in a 24-hour cycle. It has evolved as an adaptive system for living organisms in a cyclical natural environment. A regular circadian rhythm can be associated with good health, well-being, strong immunity, and high economic characteristics. The interaction between circadian rhythmicity and the physiology of farm animals is becoming an important focus of animal science. Understanding the circadian genes (CLOCK, BMAL, PER1, PER2, CRY1, CRY2) and actions regulating circadian oscillation in farm animals is important to improve management and increase economic traits. The study aimed to summarise the research on the effects of circadian rhythm genes on productivity and to highlight the importance of this topic

Principle and Application of Biophotonic Chips

Contemporarily, biophotonic chips have garnered significant attention from researchers due to their strong immunity to electromagnetic interference, high sensitivity, rapid response rate, and relatively simple structural design. They have found extensive applications in fields such as medical diagnosis, food quality and safety testing, environmental monitoring, and national security. This article provides an overview of the development history, fundamental principles, applications, and fabrication processes of biophotonic chips. The earliest development of biophotonic chips can be traced back to the end of the 20th century. The basic principle involves the interaction of light, utilizing technologies like photonic crystals, surface plasmon resonance, and fluorescence spectroscopy to achieve the detection and analysis of biological molecules. The article briefly describes the fabrication process of biophotonic chips, which involves intricate techniques such as photolithography and electron beam exposure. Furthermore, it analyzes and discusses their widespread applications in medical diagnosis, drug development, and environmental monitoring, among other fields. Lastly, the study evaluates the key challenges present in biophotonic chips and provides an analysis of their future development prospects.

Nanoparticles and Antiviral Vaccines.

Viruses have threatened human lives for decades, causing both chronic and acute infections accompanied by mild to severe symptoms. During the long journey of confrontation, humans have developed intricate immune systems to combat viral infections. In parallel, vaccines are invented and administrated to induce strong protective immunity while generating few adverse effects. With advancements in biochemistry and biophysics, different kinds of vaccines in versatile forms have been utilized to prevent virus infections, although the safety and effectiveness of these vaccines are diverse from each other. In this review, we first listed and described major pathogenic viruses and their pandemics that emerged in the past two centuries. Furthermore, we summarized the distinctive characteristics of different antiviral vaccines and adjuvants. Subsequently, in the main body, we reviewed recent advances of nanoparticles in the development of next-generation vaccines against influenza viruses, coronaviruses, HIV, hepatitis viruses, and many others. Specifically, we described applications of self-assembling protein polymers, virus-like particles, nano-carriers, and nano-adjuvants in antiviral vaccines. We also discussed the therapeutic potential of nanoparticles in developing safe and effective mucosal vaccines. Nanoparticle techniques could be promising platforms for developing broad-spectrum, preventive, or therapeutic antiviral vaccines.

Endoplasmic Reticulum-Targeting AIE Photosensitizers to Boost Immunogenic Cell Death for Immunotherapy of Bladder Carcinoma.

Bladder cancer is characterized by high rates of recurrence and multifocality. Immunogenic cell death (ICD) of cancer cells has emerged as a promising strategy to improve the immunogenicity of tumor cells for enhanced cancer immunotherapy. Although photosensitizer-based photodynamic therapy (PDT) has been validated as capable of inducing ICD in cancer cells, the photosensitizers with a sufficient ICD induction ability are still rare, and there have been few reports on the development of advanced photosensitizers to strongly evoke the ICD of bladder cancer cells for eliciting potent antitumor immune responses and eradicating bladder carcinoma in situ. In this work, we have synthesized a new kind of endoplasmic reticulum (ER)-targeting aggregation-induced emission (AIE) photosensitizer (named DPASCP-Tos), which could effectively anchor to the cellular ER and trigger focused reactive oxygen species (ROS) production within the ER, thereby boosting ICD in bladder cancer cells. Furthermore, we have demonstrated that bladder cancer cells killed by ER-targeted PDT could serve as a therapeutic cancer vaccine to elicit a strong antitumor immunity. Prophylactic vaccination of the bladder cancer cells killed by DPASCP-Tos under light irradiation promoted the maturation of dendritic cells (DCs) and the expansion of tumor antigen-specific CD8+ T cells in vivo and protected mice from subsequent in situ bladder tumor rechallenge and extended animal survival. In summary, the ER-targeted AIEgens developed here significantly amplified the ICD of bladder cells through focused ROS-based ER oxidative stress and transformed bladder cancer cells into the therapeutic vaccine to enhance immunogenicity against orthotopic bladder cancer, providing valuable insights for bladder carcinoma treatment.

Ice Identification with Error-Accumulation Enhanced Neural Dynamics in Optical Remote Sensing Images

Arctic sea ice plays an important role in Arctic-related research. Therefore, how to identify Arctic sea ice from remote sensing images with high quality in an unavoidable noise environment is an urgent challenge to be solved. In this paper, a constrained energy minimization (CEM) method is applied for Arctic sea ice identification, which only requires the target spectrum. Moreover, an error-accumulation enhanced neural dynamics (EAEND) model with strong noise immunity and high computing accuracy is proposed to aid with the CEM method for Arctic sea ice identification. With the theoretical analysis, the proposed EAEND model possesses a small steady-state error in noisy environments. Finally, compared with other existing models, the proposed EAEND model can not only complete sea ice identification in excellent fashion, but also has the advantages of high efficiency and noise immunity.

Processing Accuracy of Microcomb-Based Microwave Photonic Signal Processors for Different Input Signal Waveforms

Microwave photonic (MWP) signal processors, which process microwave signals based on photonic technologies, bring advantages intrinsic to photonics such as low loss, large processing bandwidth, and strong immunity to electromagnetic interference. Optical microcombs can offer a large number of wavelength channels and compact device footprints, which make them powerful multi-wavelength sources for MWP signal processors to realize a variety of processing functions. In this paper, we experimentally demonstrate the capability of microcomb-based MWP signal processors to handle diverse input signal waveforms. In addition, we quantify the processing accuracy for different input signal waveforms, including Gaussian, triangle, parabolic, super Gaussian, and nearly square waveforms. Finally, we analyse the factors contributing to the difference in the processing accuracy among the different input waveforms, and our theoretical analysis well elucidates the experimental results. These results provide guidance for microcomb-based MWP signal processors when processing microwave signals of various waveforms.

Assessment of the effects of prepartum anti-inflammatory therapies on type 1/type 2 immunity ratio using a rapid blood test

The objective of this study was to assess (1) the effects of prepartum administration of anti-inflammatory therapies on type 1/type 2 immunity ratio using a rapid blood test (D2Dx immunity test; Nano Discovery Inc.), and (2) correlations between rapid blood test scores and daily milk yield in Holstein dairy cows. At 14 d before the expected calving date, cows (n = 64) and heifers (n = 23) were blocked by body condition score (optimal = 3.25-3.5; high ≥3.75) and parity (nulliparous, parous), and randomly allocated to one of 3 treatment groups (1) ASA (n = 29) = receive one oral treatment with administration of acetylsalicylic acid (4 boluses; 480 grain/bolus); (2) MEL (n = 31) = receive one oral administration with meloxicam (1 mg/kg of body weight), or (3) PLC (n = 27) = receive one oral treatment with 4 gelatin capsules filled with water. Blood samples were collected weekly starting 1 wk before treatment until 3 wk after calving for assessment of type 1/type 2 immunity ratio using a rapid blood test (i.e., D2Dx immunity test). A higher D2Dx score corresponds to a higher type 1/type 2 ratio. Furthermore, blood samples were collected within 72 h before and after calving by farm personnel. Daily milk yield for the first 60 d in milk (DIM) was collected from on-farm computer records. The data were analyzed using MIXED procedure of SAS (SAS Institute Inc.) as a randomized complete block design. On average enrolled cows received treatment administration 10 d before the actual calving date (standard deviation = 5.10 d). There was a tendency for a treatment by day interaction. Cows treated with ASA had higher type 1/type 2 ratio within 3 d after calving compared with MEL and PLC cows (ASA = 0.065 ± 0.002; MEL = 0.059 ± 0.002; PLC = 0.053 ± 0.002). Similarly, ASA and MEL cows had a higher type 1/type 2 ratio at 7 ± 3 DIM compared with PLC cows (ASA = 0.062 ± 0.002; MEL = 0.064 ± 0.002; PLC = 0.056 ± 0.002). Regardless of treatment, there was an interaction between parity and day. Parous cows had higher type 1/type 2 ratios compared with nulliparous cows at 14 ± 3 d before calving and at 7 ± 3, 14 ± 3, and 21 ± 3 d after calving. Furthermore, there was a positive correlation between D2Dx scores at 14 ± 3 DIM and average daily milk yield in the first 60 DIM. These results suggest that prepartum anti-inflammatory therapies may cause an increased shift in type 1 immunity around calving. Similarly, parous cows may have an increased shift in type 1 immunity after calving. Interestingly, higher type 1/type 2 ratios may be associated with higher milk yields in the first 60 DIM. Larger studies are needed to identify associations between the D2Dx immunity test and cow health and performance, as well as to assess the applicability of these types of tests in a conventional farm setting.