Genomics of FOXP3 Enhance Treg and Maintain Immune-homeostasis

The immunologic theorem is a subject of self also non-self differentiation directly combat infections and maintains the energy of self-antigens. Immune privilege of T-cells is prevailing via crucial and supercial prospect. The T-cell promotes the function of immune replication and implant antigens. The regulatory T-cells (Treg) impart the progress of diseases also prevent immunity. X chromosome encoded FOXP3 is a regulator of segregation and immunosuppressive function. Nuclear factor FOXP3 regulated lineage-specic polarity of Treg crucially maintains immune-homeostasis. The functional inhibitions of helper T cells by FOXP3-inhibitory peptide constitute an imprint to enhance immunotherapy. The enlightenment of FOXP3 contributed to a novel concern in the experiment of suppressor T lymphocytes and mechanisms of immune homeostasis. In this exploration, I aimed to view FOXP3 functions from the FOX family in Homo sapiens and compare them with Mus musculus. Therefore, I performed a bioinformatics pipeline for the experimentation of the forkhead box P3 and their family. My nding data supported the FOX family of TF’s play a preventive strategy during the development. A ray of FOXP3 enhances Treg and maintains immunity against infections. The specic bioinformatics analysis epitomized FOXP3 as a T-cell dependent gene that can help to interpret the outcome in infection biology. of pathogenesis raises a step for new and exciting experiments mandatory to explore the feature of our biology. Target sequence retrieved from the various species-specic databases (NCBI, UniProt, KEGG, GenBank, EMBL, and DDBJ) and performed web-based application SMART for identication of particular domain in the target sequence. SWISS-MODEL is a bioinformatics web-server for comparative modelling of three-dimensional structures. This application performed for generating a 3D structure. That application routinely used in many practical applications. The SWISS-MODEL is an updated database of comparative and 3D model of organism proteome for biomedical research.

of infections divided into two types: (1) acute infections and (2) chronic infections. Acute infections commonly result from active immune reactions and chronic infections associated with suboptimal T-cell function. In this object, my focus on different infections and knowledge of the CD8 + T-cell polarity appears when the antigen eliminates via prime infection and how T-cell reactions can change and reduce chronic infections. The chronic infection effected by cytotoxic T-cells (CD8 + T-cell) of the immune process. The CD8 + T-cell reactions steadily reduce infections then cells gradually evacuate function during chronic illness. These processes enhance by helper CD4 + T-cells that control the infection and contribute a reaction of CD8 + T-cells [3,4]. Those concepts in the immune process raise a question, how and why CD4 + T-cells enhance CD8 + T-cells reaction [5][6][7]? An immunogenic function of cytokines generates by CD4 + T-cells is well-known as interleukin-21 (IL-21) that governs chronic infections. The IL-21 promotes by the process of different CD4 + T-cells subgroup i.e. T FH , extended T-helper 17 cells and LGL [8,9]. The cytokine promotes juxtapose with these subgroups and help other immune cells like large granular lymphocytes (LGL), B-cells and cytotoxic T cell for a proper response of IL-21. The IL-21R element of the gamma chain family of cytokine receptor included IL-2 (CD25), IL-7, IL-15, and IL-21 combine to improve CD8 + T-cell reactions [10][11][12]. Those groups compare to CD4 + T-cell functions for CD8 + T-cell response are associated with IL-21 as an intermediator. During this activity, the newly synthesized proteins will reduce peptide fragments even infected cells lose cytotoxic granules. The stimulation of CD8 + T-cells produces clear lymph nodes, whereas antigen appearance cells and naive Tcell interact with one another. In the lymph node, DC and CD4 + T-cells contribute to the co-stimulation is vital for con rmation of the tendency of CD8 + T-cells. Furthermore, the initial face of chronic and acute infections, Th1, Th2 and Th17, CD8+, CD4 + cells negotiate preventive immunity against illness [13,14].
The immune reaction moderate by regulatory cells like Treg, M2-type macrophages, MDSCs, TGF-β, cytokines and IL-10 innate adaptive response in immune-checkpoints that control T-cell activation. These regulatory cells in immune checkpoints enhance infections as a mechanism of immune breakthrough and become unique targets for treatment. However, the hypothesis of FOXP3 functions in Treg cells linked with MHC2 that control CD4 + T-cells also expresses a high equilibrium of IL-2 (CD25). An addition of FOXP3 function in CD4 + T-cells and IL2 appears by the process of MHC1 governs CD8 + T-cells reactions. The CD8 + T-cells can kill pathogens by reactions of perforin and granzymes, which disrupt the plasma membranes of infected cells. Also, Treg included Tr1, Th cells, CD8, CD28 and HLA-E control Tcells. The Treg activity associated with CTLA-4 and CD28 receptor potentially bound with two natural ligands such as B7-1 and B7-2 effective for Treg-suppressive capacity [15][16][17]. The CD86 sharply enhance suppression by the exercise of CD25 + and CD4 + cells. Invariance, resistance mechanisms of CD80 enhances proliferative response by reducing Treg suppression. The combined effect of B7 and B7.2 on DC regulates for improvement of unstable to a stable state with the potentiality of Treg-suppressive response. The B7-2 and B7-1 emulate functions via CD152 and CD28 on Treg that has a remarkable outcome on the repression of immune reaction. The tendency of Th cells in immune checkpoints included CTLA-4, PD-1 (CD279), TIM-3, LAG-3 and TIGIT. Furthermore, the signi cant function of PD-1 mainly expresses on T-cells that negotiate with their two ligands PD-L1 (B7-H1, CD274) and PD-L2 (B7-DC, CD273) rival a unique role of initiation and maintain peripheral tolerance mechanism. The PD-L1 and PD-L2 express APCs and infected cells release a negative signal to T-cells are called T-cell exhaustion.
Therefore, antibodies binding CTLA-4, PD-1 or PD-L1 have a dynamic e ciency to enhance immunotherapy [18][19][20][21][22][23]. The defensive mechanisms suggested the X-chromosome encoded FOXP3 transcription factor gene govern T-cells polarity and immunosuppressive function. The Treg express from FOXP3 and control immune-mediated infections. The nuclear TF of FOXP3 regulates lineage-speci c con ict of the Treg that certainly a defence of the immune-homeostasis [12]. The FOXP3 function supported as a lineage associated factor in the regulatory T-cells. The activity of Treg due to loss of FOXP3 function is vigorous and untreatable immune-mediated infections. The functional inhibition of Treg by forkhead domain associated peptides initiate an impact to promote immunotherapy [24][25][26].
Recent studies supported the FOXP3 function attain unique mechanisms in the biology of Treg and cellular-immune-homeostasis. Those reports revealed that the study of FOXP3 functions and immunologic mechanisms involved with regulatory T-cells. In this work, intense evidence of the FOX family of TF's reveals a leading function during the growth of organisms. In contrast, the X chromosome encodes FOXP3 enhances Treg and improve immunity against infections.

Results:
Structural analysis of target gene: The target sequence determined the building block of nucleotides and peptides. The predominant sequence formulated of 1296 nucleotides and 431 peptides with 83 peptides residue binding to DNA is well-known as a forkhead domain ( Table 1). The polypeptide structure demonstrated the forkhead domain is involved in DNA binding. The forkhead domain is also known as the "winged helix" domain.
The analysis of three-dimensional structure determined alpha-helices also beta-sheets link up within B-DNA as a monomer interconnects with the DNA backbone. The alpha helices pretend a thick structure that extends the third helix to the major grove. The polypeptides compose a warped anti-parallel betasheet and stochastic coil linked with the minor groove ( Fig. 1).

Genome-wide analysis:
The genome-wide analysis by the HMMER algorithm obtains 113, 87 of forkhead domains in Homo sapiens and Mus musculus, respectively (Table: 2). The BLAST2 results represent 115, 87 of the homologs FOXP3 gene in Homo sapiens and Mus musculus, respectively (Table: 2). Further analysis of Gene Ontology Annotation con rmed the sequence accuracy of FOXP3 in two different organisms (Table: 3). These standalone searches concluded the FOXP3 was present in both organisms.
Domain, motifs and chromosome location analysis: The highest hits of target (FOXP3) gene selected from Homo sapiens and Mus musculus for MSA, a multiple sequence alignment (MSA) determined the conserved forkhead domain in the two different organisms. The high consensus (90%) sequence indicated the extended FOX domain and its speci c motifs ( Fig. 2 & 3). The chromosome localization study con rmed that the FOXP3 located band Xp11.23, start 49,250,436 bp also end 49,270,477 bp (Fig. 4).

Gene expression and network analysis:
The disease state study suggested the up-regulated expression of FOXP3 in 21 of COVID-19 clinical cohorts correlated in mild/asymptomatic and mild/normal blood samples. Also, the down-regulated expression of FOXP3 in 21 COVID-19 patients correlated in asymptomatic/normal blood samples. The anatomical segments of the data of human mRNA showed the medium level FOXP3 express in tissue, circulatory system, and blood (Fig. 5). The gene network analysis determined the FOXP3 interacts with other molecules. Such as runt-related transcription factor 1, TNFRSF18 (CD357), RAR-related orphan receptor gamma, CTLA-4 (CD152), GATA binding protein 3, Interleukin-2, Interleukin-2 receptor alpha chain, Interferon-gamma (IFNγ), NFATC2, those molecular interactions govern the fundamental function of FOXP3 in the cellular process (Fig. 6).

Discussion:
The defensive immune responses against infections depend on the T-cells that can control and destroy infected cells. Activation of T-cells determined by antigen-presenting cells i.e. DCs (dendritic cells), macrophages and B & T lymphocytes [27][28][29]. In this study, I discuss the prospective peptide-based targets for the protection and elimination of infections. I justify improvement that governs protective element with quality of antigen, supplement and foundation. Also, I will elucidate adaptive peptide base protection against infectious. The leading aim of this work is the examination of peptide inhibitors protein to overcome immunosuppressive response against infections. Those infection diseases such as bacteria, viruses, fungi, parasites and tumours are the origin of acute and chronic infections. Severe infections such as a common cold, cough and fever are usually clear from patients within a week. These types of infections do not cause any risk factor. However, acute and chronic illness originated from the burden of pathogenesis. Chronic infections are persistent that causes by the ine cient immune response lead to long-lasting symptoms. Following the annual pandemic results in about 3-5 million patients affected and nearly 250,000-500,000 deaths worldwide [30][31][32][33][34][35]. Acute and chronic illness causes major health impact in human because infections have a collision on health strategies to limit or control infections. According to genome sequencing, mammals evolved a re ne-immune system to cope with all kind of illness. Precisely, the acquired immune system is most important to control various infections.
The immune reactions embrace speci c antibodies against infections that can interrupt and restrain pathogenesis. Therefore, a notable breakthrough in this work is the function and mechanisms of regulatory T-cells (Tregs) are a clear lymphocyte lineage endowed with inhibitory properties that effected catalytic reaction in the immune system. Treg (CD4 + cells) described by the function of FOXP3 (speci c forkhead box/winged-helix transcription factor 3), which is vital for the growth of organisms [36][37][38]. In this work, my nding suggested the peptide inhibitor can control the e cacy of immunity because the signal of the conserved forkhead domain appears inside the cell. Most prominently, the migration of FOXP3 nuclear translocation probably inhibits the adaptability of Treg. That process ensues FOXP3 ability to recover proliferation and enhance Treg. The FOXP3 being a homo-oligomeric element with paramount molecular substance, inconsistent polypeptides interact with different molecules such as RUNX1, TNFSF18, CTLA4, GATA-3, RORC, NFATC2, IL2, IL2RA, IFNG and IL10. So, blocking interactions between IL-10R and PD-L1 outcome of infections eliminated [39][40][41][42][43][44][45][46]. It logically assumed that the interactions of peptides and other molecules govern the ultimate function of FOXP3. Also, the interruption of participants might decline the innate reaction. The combined concepts of immunologic and molecular mechanisms contributed primary evidence of suppressive functions of FOXP3-dependent-Treg are vital for immune-homeostasis [12]. Therefore, Treg activity is dynamic for defence against various infections. Those mechanisms illustrated the elevated number of speci c CD4 + T-cells or Treg cells infected by pathogens. The peptide-base targets should elicit a robust T-cell renewal since the T-cells require reacting immediately after infections to terminate the pathogens before it causes illness or disease burden. That protective strategy evokes responses required speci c translocation is valuable. The broad and speci c Tcell reaction can control infections because T-cell response should be productive against infections without harms. The peptide-based targets can formulate antibodies also T-cell response since the peptides are synthetic, safe and secure to produce [47]. Di cult to prescribe accurate targets also prefer leading molecules that improve immunity and drive immune reactions in conscious treatment.

Conclusion Remarks:
The present nding concluded the FOX family play dominant roles during the development of organisms.
The ray of the FOXP3 gene identi ed as a T-cell dependent gene is an effective target for preventive strategy in mammals since the major T-cell dependent gene can de ne the real spectrum of infections. Besides, the fusion genes or genetic recombination may be an earlier marker for research and development. The speci c fusion genes or genetic recombination may be immunogenic and e cient in inducing IgG antibody. Therefore, the statement of pathogenesis raises a step for new and exciting experiments mandatory to explore the feature of our biology.

Materials And Methods:
Query sequence and database Target sequence retrieved from the various species-speci c databases (NCBI, UniProt, KEGG, GenBank, EMBL, and DDBJ) and performed web-based application SMART for identi cation of particular domain in the target sequence. SWISS-MODEL is a bioinformatics web-server for comparative modelling of threedimensional structures. This application performed for generating a 3D structure. That application routinely used in many practical applications. The SWISS-MODEL is an updated database of comparative and 3D model of organism proteome for biomedical research.

Genome
The draft genome sequence downloaded from genomic data in various specialized databases (Ensemble and NCBI).
Standalone tools HMMER executes multiple sequence alignments of the speci c domain as a pro le search. HMMER is a statistical algorithm that offers to make MSA of the particular domain as a pro le search is a probabilistic model called the pro le HMM. Standalone BLAST performs for homologs gene in organisms.

Annotation
The BLAST2GO performed for gene annotation is a bioinformatics and computational tool for highthroughput gene ontology annotation. The functional statistics of genes recompense via GO annotation is a glossary of the working attribute.

Domain
For examination of sustain domain in sequences. The MSA method is applicable to reckon the highest match of homologs sequences and then observe similarity, identity and differences. MSA of maximum hits of target sequences examination by web-based tool MultAlin for validation of sustain domain.

Motif
The MEME tool performs for examination of sequence-speci c motifs. MEME suite is a bioinformatics and computational web base tool to discover and validation of the sequence-speci c motifs.

Chromosome location
The chromosome location recti ed using the gene card. The gene card is a database of human provides statistics of all predicted human genes. That database is presently available for medical research like gene, protein and associated disease.

Gene networks
The genetic network is a cluster of molecules that regulate and interact with each other in the cells to command the development levels of proteins or mRNA. Some protein serves to activate genes are TF's that bind to the pioneer area and initiate the activity of other proteins is called regulatory cascades. I retrieve the STRING database for the prediction of protein-protein interaction. The STRING database contains various resources like experimental data and computational methods. The work furnished in this paper is original and communicated by the correspondent given in the manuscript. The author disclosed that the document is not concern elsewhere and not receive for evaluation by other journals.
Ethical approval: The study contains an in-silico analysis of the mammalian genome to examine the particular gene in different organisms.
Availability of data and material: The data and materials have not been deposited yet in the database. The HMMER, BLAST2, and BLAST2GO (gene ontology annotation) data are available by request or demand.

Competing interest:
The author stated that the work has no con ict of interest.

Funding:
The author did not avail of nancial assistance from any source in undertaking the present study.
Author's contributions: This research paper contains an author placed at the top of the manuscript. The author conceived the idea, experimented, analyzed data and also prepared the manuscript.      FOXP3 interacts with different TF's (Gene Network)